Validation Data Gallery

Filter: AllWBIPIHCIF/ICC

WB analysis using 10449-1-AP
MG132 treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 10449-1-AP (GADD34 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.
WB analysis of PC-3 using 10449-1-AP
WB result of GADD34 antibody (10449-1-AP, 1:500) with si-Control and si-GADD34 transfected PC-3 cells.
WB analysis of HepG2 using 10449-1-AP
HepG2 cells were subjected to SDS PAGE followed by western blot with 10449-1-AP (GADD34 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.
WB analysis of K-562 using 10449-1-AP
K-562 cells were subjected to SDS PAGE followed by western blot with 10449-1-AP (GADD34 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.
WB analysis of Jurkat using 10449-1-AP
Jurkat cells were subjected to SDS PAGE followed by western blot with 10449-1-AP (GADD34 antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.
WB analysis using 10449-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 10449-1-AP (GADD34 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.
IP experiment of NIH/3T3 using 10449-1-AP
IP result of anti-GADD34 (IP:10449-1-AP, 4ug; Detection:10449-1-AP 1:800) with NIH/3T3 cells lysate 1320 ug.
IP experiment of K-562 using 10449-1-AP
IP result of anti-GADD34 (IP:10449-1-AP, 4ug; Detection:10449-1-AP 1:800) with K-562 cells lysate 1360 ug.
IHC staining of human pancreas using 10449-1-AP
Immunohistochemical analysis of paraffin-embedded human pancreas tissue slide using 10449-1-AP (GADD34 antibody) at dilution of 1:200 (under 10x lens).
IHC staining of human pancreas using 10449-1-AP
Immunohistochemical analysis of paraffin-embedded human pancreas tissue slide using 10449-1-AP (GADD34 antibody) at dilution of 1:200 (under 40x lens).
IHC staining of human colon cancer using 10449-1-AP
Immunohistochemical analysis of paraffin-embedded human colon cancer using 10449-1-AP (GADD34 antibody) at dilution of 1:100 (under 10x lens).
IHC staining of human colon cancer using 10449-1-AP
Immunohistochemical analysis of paraffin-embedded human colon cancer using 10449-1-AP (GADD34 antibody) at dilution of 1:100 (under 40x lens).
IF Staining of U2OS using 10449-1-AP
Immunofluorescent analysis of (-20°C Ethanol) fixed MG132 treated U2OS cells using GADD34 antibody (10449-1-AP) at dilution of 1:400 and CoraLite®594-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-4).

Featured Product
KD/KO Validated

GADD34 Polyclonal antibody

GADD34 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 10449-1-AP

Publications(135)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (4)

Applications

WB, IHC, IF/ICC, IP, ELISA

Synonyms

PPP1R15A, Growth arrest and DNA damage-inducible protein GADD34, growth arrest and DNA damage-inducible protein 34

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (135)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	MG132 treated NIH/3T3 cells, Tunicamycin treated HeLa cells, PC-3 cells, K-562 cells, HepG2 cells, Jurkat cells
Positive IP detected in	K-562 cells, NIH/3T3 cells
Positive IHC detected in	human pancreas tissue, human colon cancer tissue **Note: suggested antigen retrieval with TE buffer pH 9.0; () Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0***
Positive IF/ICC detected in	MG132 treated U2OS cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:6000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:50-1:500
Immunofluorescence (IF)/ICC	IF/ICC : 1:200-1:800
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

KD/KO	See 12 publications below
WB	See 126 publications below
IHC	See 11 publications below
IF	See 10 publications below
IP	See 3 publications below
FC	See 1 publications below

Product Information

10449-1-AP targets GADD34 in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse samples.

Tested Reactivity	human, mouse
Cited Reactivity	human, mouse, rat, pig, monkey, hamster
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag0578 Product name: Recombinant human GADD34 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 307-572 aa of BC003067 Sequence: SDEEEGEVKALGAAEKDGEAECPPCIPPPSAFLKAWVYWPGEDTEEEEDEEEDEDSDSGSDEEEGEAEASSSTPATGVFLKSWVYQPGEDTEEEEDEDSDTGSAEDEREAETSASTPPASAFLKAWVYRPGEDTEEEEDEDVDSEDKEDDSEAALGEAESDPHPSHPDQRAHFRGWGYRPGKETEEEEAAEDWGEAEPCPFRVAIYVPGEKPPPPWAPPRLPLRLQRRLKRPETPTHDPDPETPLKARKVRFSEKVTVHFLAVWAG 相同性解析による交差性が予測される生物種
Full Name	protein phosphatase 1, regulatory (inhibitor) subunit 15A
Calculated molecular weight	73 kDa
Observed molecular weight	100 kDa
GenBank accession number	BC003067
Gene Symbol	GADD34
Gene ID (NCBI)	23645
RRID	AB_2168724
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	O75807
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

GADD34, also named PPP1R15A, belongs to the PPP1R15 family. GADD34 can be triggered as a direct target of activating transcription factor4 (ATF4) under ER stress, it plays a pivotal role in the recovery of cells from shut-down of translation induced by ER stress. It recruits the serine/threonine-protein phosphatase (PP1) to dephosphorylate the translation initiation factor eIF2alpha, thereby reversing the shut-off of protein synthesis initiated by stress-inducible kinases and facilitating recovery of cells from stress. GADD34 down-regulates the TGF-beta signaling pathway by promoting dephosphorylation of TGFB1 via PP1. It may also promote apoptosis by inducing TP53 phosphorylation on 'Ser-15'. Starvation-induced expression of GADD34 reduced mTOR activity and induced autophagy in wild-type mice, but not in GADD34 KO mice. Molecular weight of GADD34 is 100 kDa confirmed in GADD34 KO mice, and Proteintech's GADD34 antibody 10449-1-AP primarily recongize the 100 kDa band.

Protocols

Product Specific Protocols
IF protocol for GADD34 antibody 10449-1-AP	Download protocol
IHC protocol for GADD34 antibody 10449-1-AP	Download protocol
IP protocol for GADD34 antibody 10449-1-AP	Download protocol
WB protocol for GADD34 antibody 10449-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
	WB	Nature Stress response silencing by an E3 ligase mutated in neurodegeneration Authors - Diane L Haakonsen View Article
human	WB	Nature An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer. Authors - Soucy Teresa A TA View Article
mouse	WB,IHC	Nature Sustained translational repression by eIF2α-P mediates prion neurodegeneration. Authors - Moreno Julie A JA View Article
human	WB	Science Preventing proteostasis diseases by selective inhibition of a phosphatase regulatory subunit. Authors - Indrajit Das View Article
human	WB	Cell Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B. Authors - Agnieszka Krzyzosiak View Article
human	WB, IP	Science Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis. Authors - Pavel Tsaytler View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 10449-1-AP

WB analysis of PC-3 using 10449-1-AP

WB analysis of HepG2 using 10449-1-AP

WB analysis of K-562 using 10449-1-AP

WB analysis of Jurkat using 10449-1-AP

WB analysis using 10449-1-AP

IP experiment of NIH/3T3 using 10449-1-AP

IP experiment of K-562 using 10449-1-AP

IHC staining of human pancreas using 10449-1-AP

IHC staining of human pancreas using 10449-1-AP

IHC staining of human colon cancer using 10449-1-AP

IHC staining of human colon cancer using 10449-1-AP

IF Staining of U2OS using 10449-1-AP

GADD34 Polyclonal antibody

GADD34 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

Stress response silencing by an E3 ligase mutated in neurodegeneration

An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer.

Sustained translational repression by eIF2α-P mediates prion neurodegeneration.

Preventing proteostasis diseases by selective inhibition of a phosphatase regulatory subunit.

Target-Based Discovery of an Inhibitor of the Regulatory Phosphatase PPP1R15B.

Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis.