GADD34 Mouse Monoclonal Antibody [A4D7]
Catalog# EM1902-26
GADD34 Mouse Monoclonal Antibody [A4D7]
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WB
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IHC-P
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Human
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unconjugated
Overview
Product Name
GADD34 Mouse Monoclonal Antibody [A4D7]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Synthetic peptide within Human GADD34 aa 1-50 / 674.
Species Reactivity
Human
Validated Applications
WB, IHC-P
Target Molecular Weight
Predicted band size: 73 kDa
Positive Control
K-562 (Human chronic myelogenous leukemia cell) cell lysate, HEL (Human erythroleukemia cell) cell lysate, Caco-2 (Human colorectal adenocarcinoma cell) cell lysate, Hep G2 (Human hepatocellular carcinoma cell) cell lysate, A549 cell lysate, human prostate carcinoma tissue.
Conjugation
unconjugated
Clone Number
A4D7
RRID
Product Features
Form
Liquid
Concentration
2 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein G affinity purified.
Application Dilution
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WB
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1:5,000
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IHC-P
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1:50-1:200
Target
Function
Protein phosphatase 1 regulatory subunit 15A also known as growth arrest and DNA damage-inducible protein GADD34 is a protein that in humans is encoded by the PPP1R15A gene. The Gadd34/MyD116 gene was originally discovered as a member in a set of gadd and MyD mammalian genes encoding acidic proteins that synergistically suppress cell growth. Later on it has been characterized as a gene playing a role in ER stress-induced cell death, being a target of ATF4 that plays a role in ER-mediated cell death via promoting protein dephosphorylation of eIF2α and reversing translational inhibition. This gene is a member of a group of genes whose transcript levels are increased following stressful growth arrest conditions and treatment with DNA-damaging agents. The induction of this gene by ionizing radiation occurs in certain cell lines regardless of p53 status, and its protein response is correlated with apoptosis following ionizing radiation.
Background References
1. Song P. et. al. The regulatory protein GADD34 inhibits TRAIL-induced apoptosis via TRAF6/ERK-dependent stabilization of myeloid cell leukemia 1 in liver cancer cells. J Biol Chem. 2019 Apr
2. Goh CW. et. al. Chronic oxidative stress promotes GADD34-mediated phosphorylation of the TAR DNA-binding protein TDP-43, a modification linked to neurodegeneration. J Biol Chem. 2018 Jan
Sequence Similarity
Belongs to the PPP1R15 family.
Post-translational Modification
Phosphorylated at multiple Ser/Thr residues. Phosphorylated on tyrosine by LYN; which impairs its antiproliferative activity. Phosphorylation at Tyr-262 enhances proteasomal degradation, this position is dephosphorylated by PTPN2.; Polyubiquitinated. Exhibits a rapid proteasomal degradation with a half-life under 1 hour, ubiquitination depends on endoplasmic reticulum association.
Subcellular Location
Endoplasmic reticulum membrane, Mitochondrion outer membrane.
UNIPROT
Synonyms
Apoptosis associated protein antibody
GADD 34 antibody
Growth arrest and DNA damage inducible 34 antibody
Growth arrest and DNA damage-inducible protein GADD34 antibody
Myeloid differentiation primary response protein MyD116 homolog antibody
Ppp1r15a antibody
PR15A_HUMAN antibody
Protein phosphatase 1 regulatory (inhibitor) subunit 15A antibody
Protein phosphatase 1 regulatory subunit 15A antibody
Images
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Western blot analysis of GADD34 on different lysates with Mouse anti-GADD34 antibody (EM1902-26) at 1/5,000 dilution.
Lane 1: K-562 (Human chronic myelogenous leukemia cell) cell lysate
Lane 2: HEL (Human erythroleukemia cell) cell lysate
Lane 3: Caco-2 (Human colorectal adenocarcinoma cell) cell lysate
Lane 3: Hep G2 (Human hepatocellular carcinoma cell) cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 1 minute 34 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1902-26, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Mouse IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 73 kDa
Observed band size: 65 kDa -
☑ Knockdown (KD)
Western blot analysis of GADD34 on different lysates with Mouse anti-GADD34 antibody (EM1902-26) at 1/2,000 dilution.
Lane 1: A549-si NT cell lysate
Lane 2: A549-si GADD34 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 73 kDa
Observed band size: 65 kDa
Exposure time: 2 minutes 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1902-26) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-GADD34 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-26, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"