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Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP

Rat Monoclonal eB144

269 References
18-8817-30 18-8817-31 18-8817-33
20 µL 50 µL 200 µL
Liquid (sterile filtered) Liquid (sterile filtered) Liquid (sterile filtered)
WB, ELISA, IF, IP, Biochemical Assay, ChIP
Mouse
Rat
$66.00 /Per Item
$129.00 /Per Item
$260.00 /Per Item
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$50.00 to US & $70.00 to Canada for most products. Final costs are calculated at checkout.

Background

Mouse IgG TrueBlot® ULTRA is a unique Anti-Mouse IgG monoclonal secondary antibody. Mouse IgG TrueBlot® ULTRA enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Western blot data with Mouse IgG TrueBlot® ULTRA, simply substitute the conventional Anti-Mouse IgG blotting reagent with Mouse IgG TrueBlot® ULTRA and follow the prescribed protocol for sample preparation and immunoblotting. Mouse IgG TrueBlot® ULTRA is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. Mouse IgG TrueBlot® ULTRA preferentially detects the non-reduced form of mouse IgG (IgG1, IgG2a, IgG2b, IgG3) over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse IgG TrueBlot® ULTRA with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/immunoblot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

Product Details

Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP - 18-8817-30
Anti-Mouse IgG HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting
Rat
Peroxidase (HRP) ULTRA
Monoclonal
IgG

Target Details

Mouse
Mouse TrueBlot® ULTRA Antibody Peroxidase Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by immunoelectrophoresis resulted in a single precipitin arc against Anti-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.

Application Details

ELISA, IP, WB
Biochemical Assay, ChIP, IF  - View References
Mouse TrueBlot® ULTRA has been tested in ELISA, IP, and Western blot and may also be used for detection in immunoblotting assays that do not employ immunoprecipitation. Mouse IgG TrueBlot® ULTRA is provided as 1000X solution. To achieve best results when detecting mouse IgG1 subtypes, we recommend performing a dot blot or titration to determine the optimal dilution factor for your desired application. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user. In order to conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5mls/blot will yield enough reagent for 8 blots. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 2. Immunoprecipitate should be completely reduced. 3. BLOTTO/Milk should be used as the blocking protein for the immunoblot.

Formulation

1.0 mg/mL
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Proclin is added as an antimicrobial agent.
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol

Shipping & Handling

Wet Ice
Store at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Expiration date is six (6) months from date of receipt.
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IF, Confocal Microscopy
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(). TAM Kinases Promote Necroptosis by Regulating Oligomerization of MLKL. Mol Cell.
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(). Irradiated Human Fibroblasts as a Substitute Feeder Layer to Irradiated Mouse 3T3 for the Culture of Human Corneal Epithelial Cells: Impact on the Stability of the Transcription Factors Sp1 and NFI. Int J Mol Sci.
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(). Pink1 regulates FKBP5 interaction with AKT/PHLPP and protects neurons from neurotoxin stress induced by MPP. J Neurochem.
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(). Epigenetic modulators hydralazine and sodium valproate act synergistically in VEGI-mediated anti-angiogenesis and VEGF interference in human osteosarcoma and vascular endothelial cells. Int J Oncol.
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WB, IB, PCA
(). Acquired Resistance of EGFR-Mutated Lung Cancer to Tyrosine Kinase Inhibitor Treatment Promotes PARP Inhibitor Sensitivity. Cell Rep.
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WB, IB, PCA
(). The Myb-MuvB Complex Is Required for YAP-Dependent Transcription of Mitotic Genes. Cell Rep.
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WB, IB, PCA
(). Expression of protein disulfide isomerase A3 and its clinicopathological association in gastric cancer. Oncol Rep.
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(). Dual Sensing of Physiologic pH and Calcium by EFCAB9 Regulates Sperm Motility. Cell.
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WB, IB, PCA
(). Cand1-Mediated Adaptive Exchange Mechanism Enables Variation in F-Box Protein Expression. Molecular Cell
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WB, IB, PCA
(). Effect of human very low-density lipoproteins on cardiotrophin-like cytokine factor 1 (CLCF1) activity. Scientific Reports
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IP, Co-IP
(). The miR-106b-25 cluster mediates breast tumor initiation through activation of NOTCH1 via direct repression of NEDD4L. Oncogene
Applications
IP, Co-IP
(). ATP depletion during mitotic arrest induces mitotic slippage and APC/CCdh1-dependent cyclin B1 degradation. Experimental & Molecular Medicine
Applications
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(). IL1RAP potentiates multiple oncogenic signaling pathways in AML. Journal of Experimental Medicine
Applications
IP, Co-IP
(). Lipid-dependent deposition of alpha-synuclein and Tau on neuronal Secretogranin II-positive vesicular membranes with age. Scientific Reports
Applications
WB, IB, PCA
(). PDI Knockdown Inhibits Seizure Activity in Acute Seizure and Chronic Epilepsy Rat Models via S-Nitrosylation-Independent Thiolation on NMDA Receptor. Frontiers in Cellular Neuroscience
Applications
WB, IB, PCA
(). The sterol-responsive RNF145 E3 ubiquitin ligase mediates the degradation of HMG-CoA reductase together with gp78 and Hrd1. Elife
Applications
IP, Co-IP
(). Gankyrin Promotes Tumor-Suppressor Protein Degradation to Drive Hepatocyte Proliferation. Cellular and Molecular Gastroenterology and Hepatology
Applications
WB, IB, PCA
(). Phase transitioned nuclear Oskar promotes cell division of Drosophila primordial germ cells. Elife
Applications
WB, IB, PCA
(). Extracellular Protein Fibulin-7 and Its C-Terminal Fragment Have In Vivo Antiangiogenic Activity. Scientific Reports
Applications
WB, IB, PCA
(). Sestrin2 facilitates glutamine-dependent transcription of PGC-1α and survival of liver cancer cells under glucose limitation. FEBS J.
Applications
WB, IB, PCA; IP, Co-IP
(). lncRNA Epigenetic Landscape Analysis Identifies EPIC1 as an Oncogenic lncRNA that Interacts with MYC and Promotes Cell-Cycle Progression in Cancer. Cancer Cell.
Applications
WB, IB, PCA
(). Modulation of Protein-Interaction States through the Cell Cycle. Cell.
Applications
IP, Co-IP
(). PELI1 Selectively Targets Kinase-Active RIP3 for Ubiquitylation-Dependent Proteasomal Degradation. Mol Cell.
Applications
WB, IB, PCA; IP, Co-IP
(). 53BP1 cooperation with the REV7-shieldin complex underpins DNA structure-specific NHEJ. Nature.
Applications
WB, IB, PCA
(). MEF2B Instructs Germinal Center Development and Acts as an Oncogene in B Cell Lymphomagenesis. Cancer Cell.
Applications
WB, IB, PCA
(). The interaction between IKKα and LC3 promotes type I interferon production through the TLR9-containing LAPosome. Sci Signal.
Applications
WB, IB, PCA
(). Active N6-Methyladenine Demethylation by DMAD Regulates Gene Expression by Coordinating with Polycomb Protein in Neurons. Mol Cell.
Applications
WB, IB, PCA
(). Flap endonuclease 1 is involved in cccDNA formation in the hepatitis B virus. PLoS Pathog.
Applications
WB, IB, PCA
(). Expression of Human Cytomegalovirus IE1 Leads to Accumulation of Mono-SUMOylated PML That Is Protected from Degradation by Herpes Simplex Virus 1 ICP0. J Virol.
Applications
WB, IB, PCA
(). NONO Detects the Nuclear HIV Capsid to Promote cGAS-Mediated Innate Immune Activation. Cell. 2018;175(2):488-501.e22. doi:10.1016/j.cell.2018.08.062 Cell.
Applications
WB, IB, PCA
(). The RNA-binding protein PTBP1 is necessary for B cell selection in germinal centers. Nat Immunol.
Applications
WB, IB, PCA
(). Original Ligand for LTβR Is LIGHT: Insight into Evolution of the LT/LTβR System. J Immunol.
Applications
WB, IB, PCA; In Vitro Binding Assay
(). The multi-subunit GID/CTLH E3 ubiquitin ligase promotes cell proliferation and targets the transcription factor Hbp1 for degradation. Elife.
Applications
WB, IB, PCA
(). Identification of hepatic protein-protein interaction targets for betaine homocysteine S-methyltransferase. PLoS One.
Applications
WB, IB, PCA
(). Inhibition of TRAF6 ubiquitin-ligase activity by PRDX1 leads to inhibition of NFKB activation and autophagy activation. Autophagy.
Applications
WB, IB, PCA
(). Elucidating the mitochondrial proteome of Toxoplasma gondii reveals the presence of a divergent cytochrome c oxidase. Elife.
Applications
WB, IB, PCA
(). Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis. Nat Commun.
Applications
WB, IB, PCA
(). Protein Phosphotyrosine Phosphatase 1B (PTP1B) in Calpain-dependent Feedback Regulation of Vascular Endothelial Growth Factor Receptor (VEGFR2) in Endothelial Cells: IMPLICATIONS IN VEGF-DEPENDENT ANGIOGENESIS AND DIABETIC WOUND HEALING. Journal of Biological Chemistry
Applications
IP, Co-IP
(). A Cytoplasmic RNA Virus Alters the Function of the Cell Splicing Protein SRSF2. Journal of Virology
Applications
IP, Co-IP
(). Interference of the complex between NCS-1 and Ric8a with phenothiazines regulates synaptic function and is an approach for fragile X syndrome. Proc. Natl. Acad. Sci. U.S.A
Applications
IP, Co-IP; WB, IB, PCA
(). An interaction domain in human SAMD9 is essential for myxoma virus host-range determinant M062 antagonism of host anti-viral function. Virology
Applications
IP, Co-IP; WB, IB, PCA
(). NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma. Oncogene
Applications
IP, Co-IP; WB, IB, PCA
(). Human mitochondrial cytochrome c oxidase assembly factor COX18 acts transiently as a membrane insertase within the subunit 2 maturation module. Journal of Biological Chemistry
Applications
IP, Co-IP; WB, IB, PCA
(). An FAK-YAP-mTOR Signaling Axis Regulates Stem Cell-Based Tissue Renewal in Mice. Cell Stem Cell
Applications
IF, Confocal Microscopy; WB, IB, PCA
(). RNA Binding Protein CUGBP1 Inhibits Liver Cancer in a Phosphorylation-Dependent Manner. Molecular and Cellular Biology
Applications
WB, IB, PCA
(). BAP1 regulates IP3R3-mediated Ca2+ flux to mitochondria suppressing cell transformation. Nature
Applications
WB, IB, PCA
(). Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis. Nature Communications
Applications
IP, Co-IP
(). Centriolar Satellites Control GABARAP Ubiquitination and GABARAP-Mediated Autophagy. Current Biology
Applications
IP, Co-IP
(). A Brucella Type IV Effector Targets the COG Tethering Complex to Remodel Host Secretory Traffic and Promote Intracellular Replication. Cell Host Microbe
Applications
WB, IB, PCA
(). A Brucella Type IV Effector Targets the COG Tethering Complex to Remodel Host Secretory Traffic and Promote Intracellular Replication. Cell Host Microbe
Applications
IF, Confocal Microscopy
(). POPX2 phosphatase regulates apoptosis through the TAK1-IKK-NF-κB pathway. Cell Death & Disease
Applications
WB, IB, PCA
(). Blue light and CO2 signals converge to regulate light-induced stomatal opening. Nature Communications
Applications
IP, Co-IP; WB, IB, PCA
(). Mechanistic Insights into Autoinhibition of the Oncogenic Chromatin Remodeler ALC1. Molecular Cell
Applications
IP, Co-IP
(). Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nature Communications
Applications
IP, Co-IP; WB, IB, PCA
(). Zinc finger protein ZPR9 functions as an activator of AMPK-related serine/threonine kinase MPK38/MELK involved in ASK1/TGF-β/p53 signaling pathways. Scientific Reports
Applications
IP, Co-IP; WB, IB, PCA
(). Angiotensin II activates CaV1.2 Ca2+channels through β-arrestin2 and casein kinase 2 in mouse immature cardiomyocytes. The Journal of Physiology
Applications
WB, IB, PCA
(). PrimPol is required for replication reinitiation after mtDNA damage. Proc Natl Acad Sci U S A.
Applications
WB, IB, PCA
(). Composition and Regulation of the Cellular Repertoire of SCF Ubiquitin Ligases. Cell.
Applications
WB, IB, PCA
(). SFPQ associates to LSD1 and regulates the migration of newborn pyramidal neurons in the developing cerebral cortex. Int J Dev Neurosci.
Applications
WB, IB, PCA
(). The Mammalian Ribo-interactome Reveals Ribosome Functional Diversity and Heterogeneity. Cell.
Applications
WB, IB, PCA
(). Human cytomegalovirus-encoded viral cyclin-dependent kinase (v-CDK) UL97 phosphorylates and inactivates the retinoblastoma protein-related p107 and p130 proteins. J Biol Chem.
Applications
WB, IB, PCA
(). Phosphorylation of transcriptional regulators in the retinoblastoma protein pathway by UL97, the viral cyclin-dependent kinase encoded by human cytomegalovirus. Virology.
Applications
WB, IB, PCA
(). A Short cis-Acting Motif in the M112-113 Promoter Region Is Essential for IE3 To Activate M112-113 Gene Expression and Is Important for Murine Cytomegalovirus Replication. J Virol.
Applications
WB, IB, PCA
(). Contribution of JAK2 mutations to T-cell lymphoblastic lymphoma development. Leukemia
Applications
IP, Co-IP
(). The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC. Journal of Virology
Applications
WB, IB, PCA
(). The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC. Journal of Virology
Applications
WB, IB, PCA; IP, Co-IP
(). Characterization of RyDEN (C19orf66) as an Interferon-Stimulated Cellular Inhibitor against Dengue Virus Replication. PLOS Pathogens
Applications
WB, IB, PCA
(). Leucine zipper motif in RRS1 is crucial for the regulation of Arabidopsis dual resistance protein complex RPS4/RRS1. Scientific Reports
Applications
IP, Co-IP; WB, IB, PCA
(). NBR1 enables autophagy-dependent focal adhesion turnover. Journal of Cell Biology
Applications
IP, Co-IP
(). Oral administration of Aloe vera gel powder prevents UVB-induced decrease in skin elasticity via suppression of overexpression of MMPs in hairless mice. Bioscience, Biotechnology, and Biochemistry
Applications
WB, IB, PCA
(). Direct Demonstration That Loop1 of Scap Binds to Loop7: A CRUCIAL EVENT IN CHOLESTEROL HOMEOSTASIS. Journal of Biological Chemistry
Applications
WB, IB, PCA
(). Cyclic Nucleotide Control of Microtubule Dynamics for Axon Guidance. The Journal of Neuroscience
Applications
WB, IB, PCA
(). Complement system activation contributes to the ependymal damage induced by microbial neuraminidase. Journal of Neuroinflammation
Applications
WB, IB, PCA
(). RNA Exosome Complex Regulates Stability of the Hepatitis B Virus X-mRNA Transcript in a Non-stop-mediated (NSD) RNA Quality Control Mechanism. Journal of Biological Chemistry
Applications
WB, IB, PCA
(). SMG7 is a critical regulator of p53 stability and function in DNA damage stress response. Cell Discovery
Applications
WB, IB, PCA
(). Cereblon negatively regulates TLR4 signaling through the attenuation of ubiquitination of TRAF6. Cell Death & Disease
Applications
WB, IB, PCA
(). Suppressor of Cytokine Signaling (SOCS)1 Regulates Interleukin-4 (IL-4)-activated Insulin Receptor Substrate (IRS)-2 Tyrosine Phosphorylation in Monocytes and Macrophages via the Proteasome. Journal of Biological Chemistry
Applications
IP, Co-IP
(). Eph-mediated tyrosine phosphorylation of citron kinase controls abscission. Journal of Cell Biology
Applications
IP, Co-IP; WB, IB, PCA
(). Rac1-mediated cytoskeleton rearrangements induced by intersectin-1s deficiency promotes lung cancer cell proliferation, migration and metastasis. Molecular Cancer
Applications
IP, Co-IP; WB, IB, PCA
(). Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila. Molecular Systems Biology
Applications
WB, IB, PCA
(). Activation of the thrombopoietin receptor by mutant calreticulin in CALR-mutant myeloproliferative neoplasms. Blood.
Applications
WB, IB, PCA
(). TRIP13 promotes error-prone nonhomologous end joining and induces chemoresistance in head and neck cancer. Nat Commun.
Applications
WB, IB, PCA
(). Loss of the antioxidant enzyme CuZnSOD (Sod1) mimics an age-related increase in absolute mitochondrial DNA copy number in the skeletal muscle. Age (Dordr).
Applications
WB, IB, PCA
(). The Oncogene PDRG1 Is an Interaction Target of Methionine Adenosyltransferases. PLoS One.
Applications
WB, IB, PCA
(). A multi-stage process including transient polyploidization and EMT precedes the emergence of chemoresistent ovarian carcinoma cells with a dedifferentiated and pro-inflammatory secretory phenotype. Oncotarget
Applications
WB, IB, PCA
(). Myotubularin-related proteins 3 and 4 interact with polo-like kinase 1 and centrosomal protein of 55 kDa to ensure proper abscission. Molecular & Cellular Proteomics
Applications
IP, Co-IP
(). A monoclonal antibody that tracks endospore formation in the microsporidium Nosema bombycis. PLOS One
Applications
WB, IB, PCA
(). Ebola Virus Glycoprotein Promotes Enhanced Viral Egress by Preventing Ebola VP40 From Associating With the Host Restriction Factor BST2/Tetherin. The Journal of Infectious Diseases
Applications
IP, Co-IP; WB, IB, PCA
(). Lipopolysaccharide Induces Alveolar Macrophage Necrosis via CD14 and the P2X7 Receptor Leading to Interleukin-1α Release. Immunity
Applications
IP, Co-IP
(). TRAF1 Coordinates Polyubiquitin Signaling to Enhance Epstein-Barr Virus LMP1-Mediated Growth and Survival Pathway Activation. PLOS Pathogens
Applications
WB, IB, PCA
(). Hypertension-causing Mutations in Cullin3 Protein Impair RhoA Protein Ubiquitination and Augment the Association with Substrate Adaptors. Journal of Biological Chemistry
Applications
IP, Co-IP; WB, IB, PCA
(). Differential Effects of Hepatocyte Nuclear Factor 4α Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. Molecular and Cellular Biology
Applications
ChIP; WB, IB, PCA
(). Loss of p53 enhances the function of the endoplasmic reticulum through activation of the IRE1α/XBP1 pathway. Oncotarget
Applications
WB, IB, PCA
(). MAZR and Runx Factors Synergistically Repress ThPOK during CD8+ T Cell Lineage Development. Journal of Immunology
Applications
IP, Co-IP; WB, IB, PCA
(). MAZR and Runx Factors Synergistically Repress ThPOK during CD8+ T Cell Lineage Development. Journal of Immunology
Applications
IP, Co-IP
(). Expression of human carcinoembryonic antigen-related cell adhesion molecule 6 and alveolar progenitor cells in normal and injured lungs of transgenic mice. Physiological Reports
Applications
WB, IB, PCA
(). TGF-β suppression of HBV RNA through AID-dependent recruitment of an RNA exosome complex. PLoS Pathog.
Applications
WB, IB, PCA
(). Silencing of PMEPA1 accelerates the growth of prostate cancer cells through AR, NEDD4 and PTEN. Oncotarget.
Applications
WB, IB, PCA
(). Molecular Determinants for the Inactivation of the Retinoblastoma Tumor Suppressor by the Viral Cyclin-dependent Kinase UL97. J Biol Chem.
Applications
WB, IB, PCA
(). Role of extracellular matrix in development of skeletal muscle and postmortem aging of meat. Meat Sci.
Applications
WB, IB, PCA
(). The conserved Trp114 residue of thioredoxin reductase 1 has a redox sensor-like function triggering oligomerization and crosslinking upon oxidative stress related to cell death. Cell Death Dis.
Applications
WB, IB, PCA
(). Differential regulation of S-region hypermutation and class switch recombination by noncanonical functions of uracil DNA glycosylase. Proc Natl Acad Sci USA
Applications
ChIP; IP, Co-IP
(). The nuclear form of glutathione peroxidase 4 colocalizes and directly interacts with protamines in the nuclear matrix during mouse sperm chromatin assembly. Spermatogenesis.
Applications
WB, IB, PCA
(). OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts. PLoS One.
Applications
WB, IB, PCA
(). Specificity of anti-tau antibodies when analyzing mice models of Alzheimer's disease: problems and solutions. PLoS One.
Applications
WB, IB, PCA
(). The COP II adaptor protein TMED7 is required to initiate and mediate the delivery of TLR4 to the plasma membrane. Sci Signal.
Applications
WB, IB, PCA
(). Lipoprotein lipase (LPL) is associated with neurite pathology and its levels are markedly reduced in the dentate gyrus of Alzheimer's disease brains. The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
Applications
WB, IB, PCA
(). RAB8B is required for activity and caveolar endocytosis of LRP6. Cell Rep.
Applications
WB, IB, PCA
(). Sfrp5 modulates both Wnt and BMP signaling and regulates gastrointestinal organogenesis [corrected] in the zebrafish, Danio rerio. PLoS One.
Applications
WB, IB, PCA
(). Aerobic exercise + weight loss decreases skeletal muscle myostatin expression and improves insulin sensitivity in older adults. Obesity (Silver Spring).
Applications
WB, IB, PCA
(). SARM1 and TRAF6 bind to and stabilize PINK1 on depolarized mitochondria. Mol Biol Cell.
Applications
WB, IB, PCA
(). High-content analysis of proapoptotic EphA4 dependence receptor functions using small-molecule libraries. J Biolmol Screen
Applications
WB, IB, PCA
(). MPS1/Mph1 phosphorylates the kinetochore protein KNL1/Spc7 to recruit SAC components. Nat Cel Biol
Applications
WB, IB, PCA
(). Host genetics and Chlamydia disease: prediction and validation of disease severity mechanisms. PLoS One.
Applications
IP, Co-IP
(). Leucine zipper domain is required for Kaposi sarcoma-associated herpesvirus (KSHV) K-bZIP protein to interact with histone deacetylase and is important for KSHV replication. J Biol Chem.
Applications
WB, IB, PCA
(). Growth differentiation factor-15 deficiency inhibits atherosclerosis progression by regulating interleukin-6-dependent inflammatory response to vascular injury. J Am Heart Assoc.
Applications
WB, IB, PCA
(). H2B homology region of major immediate-early protein 1 is essential for murine cytomegalovirus to disrupt nuclear domain 10, but is not important for viral replication in cell culture. J Gen Virol.
Applications
WB, IB, PCA
(). c-Jun N-terminal kinase phosphorylates DCP1a to control formation of P bodies. J Cell Biol.
Applications
WB, IB, PCA
(). Ins(1,4,5)P3 receptor-mediated Ca2+ signaling and autophagy induction are interrelated. Autophagy.
Applications
WB, IB, PCA
(). A genome-wide profiling of the humoral immune response to Chlamydia trachomatis infection reveals vaccine candidate antigens expressed in humans. J Immunol.
Applications
WB, IB, PCA
(). Posttranslational regulation of membrane type 1-matrix metalloproteinase (MT1-MMP) in mouse PTEN null prostate cancer cells: Enhanced surface expression and differential O-glycosylation of MT1-MMP. Biochim Biophys Acta.
Applications
WB, IB, PCA
(). Noble metal targeting of thioredoxin reductase--covalent complexes with thioredoxin and thioredoxin-related protein of 14 kDa triggered by cisplatin. Free Radic Biol Med.
Applications
WB, IB, PCA
(). Tautomycetin suppresses the TNFα/NF-κB pathway via inhibition of IKK activation. Int J Oncol
Applications
IP, Co-IP
(). A feedback loop between the androgen receptor and a NEDD4-binding protein, PMEPA1, in prostate cancer cells. J Biol Chem.
Applications
WB, IB, PCA

What is Rockland’s TrueBlot® product line?

Rockland’s TrueBlot® product line is designed to solve common experimental problems when performing immunoprecipitation/co-immunoprecipitation experiments and Western blots of immunoprecipitated samples (IP–Western blot). The product line consists of TrueBlot® monoclonal secondary antibodies and TrueBlot® IP beads.

What is the advantage of TrueBlot® secondaries over regular secondary antibodies?

TrueBlot® antibodies are specific to the whole IgG molecule and will not bind heavy or light chains. This is useful for binding target proteins with an expected MW near 25 kDa (light chains) and 50 kDa (heavy chains).

Why does it appear that the TrueBlot® antibody is binding heavy/light chains in my sample?

The sample must be fully reduced to eliminate cross-reactivity with heavy and light chains. Any reactivity to heavy and light chains could be due to incomplete reduction of your sample. Please be sure to optimize reducing conditions for your sample type.

How can I ensure my samples are fully reduced?

Samples can be fully reduced by heating at 90-100°C for 10 minutes in sample buffer containing reducing agent (for example, DTT to a final concentration of 50 mM or add β-Mercaptoethanol to a final concentration of 2% (v/v)).

With which species/isotypes are TrueBlot® secondary antibodies reactive?

TrueBlot® secondary antibodies are reactive with the IgG of their respective species. For example, if using a primary antibody raised in mouse (mouse host), use TrueBlot® anti-mouse Ig secondary antibodies for detection.

Do TrueBlot® secondary antibodies detect IgM?

TrueBlot® secondary antibodies have not been shown to detect IgM.

Do I need to preclear my lysate before the immunoprecipitation step?

Samples that have many other proteins present (such as lysates) may require preclearing to prevent interference in later IP and WB procedures. Recombinant protein samples require pre-clearing more often than serum samples.

How can I enrich/concentrate my lysate for a low expression of a protein of interest?

The immunoprecipitation procedure can be repeated several times to yield a more concentrated protein solution.

What is the recommended lysis buffer for TrueBlot® products?

The optimal lysis buffer will depend on your sample type. See protocol for buffer recommendations. Generally, 1X RIPA is used to lyse samples.

My target protein has the same MW as a heavy/light chain. How can I be sure that the band is the target protein and not a heavy/light chain?

Be sure to include a positive control for the primary antibody in your experiment and check that the sample is fully reduced.

What is the average size of TrueBlot® magnetic/agarose beads?

The beads are roughly 0.5 μm in diameter.

Why can’t I use Protein A or Protein G beads for the IP step in IP–Western blot when using rabbit species?

Using Protein A or Protein G beads with rabbit species results in contaminated bands. For best results when using rabbit species, use Rockland’s TrueBlot® Rabbit Ig IP beads, which are available in either a magnetic or agarose format. Please see individual TrueBlot® Rabbit product pages for additional details.

Can I use Protein A or Protein G beads for the IP step in IP–Western blot when using mouse, goat, or sheep species?

Yes. If you are using Protein A or Protein G beads with mouse, goat, or sheep species, we recommend the following: Determine the compatibility of Protein A or Protein G with your species and subtype, do not use excessive amounts of slurry, and choose the appropriate elution conditions. In some instances, harsher elution conditions can cause stripping of the subunits of Protein A or Protein G from the bead support and result in non-specific bands. For best results, we recommend using TrueBlot® Ig IP beads, which are available in either agarose or magnetic format.

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This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.