GADD34 Polyclonal antibody

• Featured Product
• KD/KO Validated

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

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (4)

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

Publications

122

Reviews (4)

Cat no : 10449-1-AP

Synonyms

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

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Validation Data Gallery

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  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.

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  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.

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  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.

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  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.

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  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.

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  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.

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  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).

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  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).

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  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).

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  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).

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  IF Staining of PC-3 using 10449-1-AP

  Immunofluorescent analysis of PC-3 cells using 10449-1-AP (GADD34 antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Rabbit IgG(H+L).

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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: PC-3 cells

Recommended 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 : 1:20-1:200

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 117 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: GADD34 fusion protein Ag0578
• 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: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

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.

Publications

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

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Ana (Verified Customer) (03-11-2021)

GADD34 in mouse muscle. 10 ug of protein. BSA blocking. 1:1000 primary ab dilution O/N incubation 4ºC.1:5000 secondary ab HRP conjugated.

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Mouse Muscle

GADD34 Antibody Western Blot validation (1:1000 dilution) in Mouse Muscle (Cat no:10449-1-AP)

FH

Maria (Verified Customer) (02-10-2021)

GADD34 in human primary fibroblasts. UT and TN treated. PFA 4% 20 min. Blocking: PBST (0.1% Triton) 10% Goat Serum 1h RT. Primary incubation: 16h 4ºC 1:200 in PBST 10% GS. Secondary: 2h RT 1:500 Goat anti-rabbit Alexa fluor 555 in PBST 10% GS.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:200
• Cell Tissue Type: Human primary fibroblasts

GADD34 Antibody Immunofluorescence validation (1:200 dilution) in Human primary fibroblasts (Cat no:10449-1-AP)

FH

Daniel (Verified Customer) (10-01-2019)

Cytoplasmic staining of the tubules of wild-type kidneys.

• Applications: Immunohistochemistry,
• Primary Antibody Dilution: 1:100
• Cell Tissue Type: Mouse kidney

GADD34 Antibody Immunohistochemistry, validation (1:100 dilution) in Mouse kidney (Cat no:10449-1-AP)

FH

Stefan (Verified Customer) (07-31-2019)

See publication:Dalton LE, Clarke HJ, Knight J, Lawson MH, Wason J, Lomas DA, Howat WJ, Rintoul RC, Rassl DM, Marciniak SJ. (2013). The endoplasmic reticulum stress marker CHOP predicts survival in malignant mesothelioma. Br J Cancer. 2013 Apr 2;108(6):1340-7. doi: 10.1038/bjc.2013.66. Epub 2013 Feb 14.https://www.nature.com/articles/bjc201366

• Applications: Western Blot, Immunohistochemistry,
• Primary Antibody Dilution: WB 1:1000, IHC 1:100
• Cell Tissue Type: HCT116 human colon carcinoma

GADD34 Antibody Western Blot,Immunohistochemistry, validation (WB 1:1000, IHC 1:100 dilution) in HCT116 human colon carcinoma (Cat no:10449-1-AP)

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