GADD153/CHOP Antibody (9C8)

Images

 
Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - Ethanol feeding increases CHOP expression. Image from verified customer review.
Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335] - FFPE tissue section of mouse brain using 1:100 dilution of Calreticulin antibody. The signal was developed using HRP-DAB based detection method ...read more
Biological Strategies: Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - GADD153/CHOP expression in HeLa cells treated with 2.5 ug/mL tunicamycin for 4 hours (Lane 1) and untreated (Lane 2).
Biological Strategies: Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - Analysis of endogenous CHOP/GADD153 from primary human fibroblasts using NB600-1335. Lane 1: Untreated cells, Lane 2: Cells treated ...read more
Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335] - Analysis of CHOP in rat heart tissue lysate. Image courtesy of product review submitted by Lee Hsiao-Wei.
Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335] - FFPE tissue section of mouse brain using 1:100 dilution of Calreticulin antibody. The signal was developed using HRP-DAB based detection method ...read more
Simple Western: GADD153/CHOP Antibody (9C8) [NB600-1335] - Image shows a specific band for CHOP/GADD153 in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, PmSpecies Glossary
Applications WB, Simple Western, GS, ICC/IF, IHC, IHC-P, IP
Clone
9C8
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1 mg/ml
Validated by:
   

Biological Strategies

     

GADD153/CHOP Antibody (9C8) Summary

Immunogen
Full length mouse CHOP/GADD153 [Swiss-Prot# P35639]
Epitope
The N-terminal region of CHOP/GADD153.
Localization
Nuclear
Marker
ER Stress Marker
Isotype
IgG2b Kappa
Clonality
Monoclonal
Host
Mouse
Gene
DDIT3
Purity
Protein A purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot
  • Simple Western 1:250
  • Gel Super Shift Assays
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 1:10 - 1:500
Application Notes
In Western blot a band can be seen at approx. 29 kDa. Gel Super Shift Assays was reported in scientific literature.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
29 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
GADD153/CHOP Lysate (NBP2-66219)
Reviewed Applications
Read 3 Reviews rated 4.3
using
NB600-1335 in the following applications:

Publications
Read Publications using
NB600-1335 in the following applications:

  • GS
    2 publications
  • 3 publications
  • 2 publications
  • IP
    4 publications
  • WB
    13 publications

Reactivity Notes

Human, mouse, rat and primate.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tris-Glycine, 0.15 M NaCl
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Purity
Protein A purified

Alternate Names for GADD153/CHOP Antibody (9C8)

  • C/EBP-homologous protein 10
  • C/EBP-homologous protein
  • CCAAT/enhancer-binding protein homologous protein
  • CEBP zeta
  • CHOP
  • CHOP10 CEBPZ
  • CHOP10
  • CHOP-10
  • CHOPC/EBP zeta
  • DDIT3
  • DDIT-3
  • DNA damage-inducible transcript 3 protein
  • DNA-damage-inducible transcript 3
  • GADD153
  • Growth arrest and DNA damage-inducible protein GADD153

Background

GADD153, also known as CHOP or DDIT3, is as a growth arrest and DNA damage-inducible gene that encodes a C/EBP-related nuclear protein. This protein functions as a transcriptional inhibitor by forming heterodimers with C/EBP (CCAAT/enhancer-binding protein) and LAP (liver activator protein), and preventing their DNA binding activity. Under normal growth conditions GADD153 is dormant; however it is highly unregulated during cellular stress caused by toxins, metabolic inhibitors and nutrient deprivation. GADD153 is present in the cytosol under nonstressed conditions and then accumulates in the nucleus once it is induced by stress.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for GADD153/CHOP Antibody (NB600-1335)(20)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Primate.

We have publications tested in 5 applications: GS, ICC/IF, IHC-P, IP, WB.


Filter By Application
GS
(2)
ICC/IF
(3)
IHC-P
(2)
IP
(4)
WB
(13)
All Applications
Filter By Species
Human
(8)
Mouse
(9)
Rat
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 20. Show All 20 Publications.
Publications using NB600-1335 Applications Species
Guo J, Cui Y, Liu Q, Yang Y. Piperine ameliorates SCA17 neuropathology by reducing ER stress. Mol Neurodegener. 2018 Jan 30 [PMID: 29378605] (WB, Rat) WB Rat
Verma S, Pillay P, Naicker T et al. Placental hypoxia inducible factor -1a & CHOP immuno-histochemical expression relative to maternal circulatory syncytiotrophoblast micro-vesicles in preeclamptic and normotensive pregnancies European Journal of Obstetrics & Gynecology and Reproductive Biology. 2017 Nov 07 [PMID: 29127866] (Human) Human
Jain A, Zode G, Kasetti RB et al. CRISPR-Cas9-based treatment of myocilin-associated glaucoma. Proc. Natl. Acad. Sci. U.S.A. 2017 Oct 17 [PMID: 28973933] (WB, ICC/IF, Human) WB, ICC/IF Human
Zhang W, Zhong W, Sun Q et al. Hepatic overproduction of 13- HODE due to ALOX15 upregulation contributes to alcohol-induced liver injury in mice. Sci Rep 2017 Aug 21 [PMID: 28827690] (WB, Mouse) WB Mouse
Kilaparty SP, Agarwal R, Singh P et al. Endoplasmic reticulum stress-induced apoptosis accompanies enhanced expression of multiple inositol polyphosphate phosphatase 1 (Minpp1): a possible role for Minpp1 in cellular stress response. Cell Stress Chaperones. 2016 Apr 02 [PMID: 27038811] (WB, Mouse)

Details:
Mouse monoclonal CHOP antibody was used for WB analysis of lysates from MC3T3-E1 cells which were incubated or not with ER stress inducers brefeldin A and thapsigargin.
WB Mouse
Ying R, Wang XQ, Yang Y et al. Hydrogen sulfide suppresses endoplasmic reticulum stress-induced endothelial-to-mesenchymal transition through Src pathway. Life Sci. 2016 Jan 01 [PMID: 26656263] (WB, Human) WB Human
Djelti F, Braudeau J, Hudry E et al. CYP46A1 inhibition, brain cholesterol accumulation and neurodegeneration pave the way for Alzheimer's disease Brain 2015 Aug 01 [PMID: 26141492] (IHC-P, Mouse)

Details:
CHOP antibody was used for IHC-P on brain tissues from wild-type C57Bl/6 and 5-month-old APP23 transgenic mice (Thy1-hAPPswe) that were subjected to injection of AAV-scramble or AAV-shCYP46A1 vectors. The IHC-P assay implicated 4% paraformaldehyde fixation at 4C for 24 hours, 6 um paraffin sections, sodium citrate based heat induced antigen retrieval, permeablization in PBS/0.1% Triton X 100, use of primary at 1:500 dilution with ON 4C incubation, detection using fluorescence labelled secondary (Figure 3A).
IHC-P Mouse
Sun Q, Zhong W, Zhang W et al. Zinc deficiency mediates alcohol-induced apoptotic cell death in the liver of rats through activating ER and mitochondrial cell death pathways. Am J Physiol Gastrointest Liver Physiol. 2015 May 01 [PMID: 25767260] (WB, Mouse) WB Mouse
Lee YJ, Ha YJ, Na Kang Y et al. The Autophagy-Related Marker LC3 Can Predict Prognosis in Human Hepatocellular Carcinoma. PLoS One. 2013 Nov 25 [PMID: 24282606] (IHC-P, Human) IHC-P Human
Lin CJ, Lee CC, Shih YL et al. Inhibition of mitochondria- and endoplasmic reticulum stress-mediated autophagy augments temozolomide-induced apoptosis in glioma cells. PLoS One 2012 [PMID: 22745676] (WB, Human) WB Human
Show All 20 Publications.

Reviews for GADD153/CHOP Antibody (NB600-1335) (3) 4.33

Average Rating: 4.3
(Based on 3 reviews)
We have 3 reviews tested in 2 species: Human, Rat.

Reviews using NB600-1335:
Filter by Applications
WB
(3)
All Applications
Filter by Species
Human
(1)
Rat
(2)
All Species
Images Ratings Applications Species Date Details
Western Blot GADD153/CHOP NB600-1335
Enlarge
reviewed by:
Anonymous
WB Human 12/15/2017
View

Summary

ApplicationWestern Blot
Sample Testedhuman liver and primary hepatocytes
SpeciesHuman
LotA-5

Comments

CommentsName: Anti-CHOP antibody (NB600-1335)
Catalog #: Anti-CHOP antibody (NB600-1335)
Lot Number: Anti-CHOP antibody (NB600-1335, Lot # A-5)
PO/Order Number: Click here to enter text..
WB Image Description (Please provide labels for all lanes): lane 1: human liver; lane 2: human primary hepatocytes
Sample Information:
     Cell Line or Tissue: human liver and primary hepatocytes
     Species: human
     Treatment: No treatment
Lysate Preparation:
     Date of lysate preparation: December 9, 2017
     Lysis buffer used: 1X lysis buffer from Cell Signaling by adding PMSF
Reducing agent: beta-mercaptoethanol, DTT
If boiled (temperature/time): Yes
Controls:
     Positive Control: No
     Negative Control: No
     Loading Control (please attach additional images if applicable): No
Protein Amount Loaded per lane: 20 ug
Antibody Storage Conditions: -20℃
Electrophoresis:
Gel Percentage: 10%
Electrophoresis Conditions: Tris-Glycine-SDS at room temperature
     Voltage: 120V
     Time: 2 hours
Membrane Transfer:
     Method (Submersion/Semi-dry): wet transfer
     Membrane Type (PVDF/Nitrocellulose): Nitrocellulose
     Time: 2 hours
     Voltage: 100V
Blocking:
     Blocking Solution: 5% milk in 1X TBST
     Time: 1 hour at room temperature
Primary Antibody:
     Dilution: 1/1000
     Diluent Buffer: 2.5% BSA
     Incubation Time: overnight
     Incubation Temperature: 4℃
Washing Conditions:
     Wash Solution: 1X TBST
     Time and Repetitions: 5 min each for 3 times
Secondary Antibody
Manufacturer and Catalog #: Promega, W402B, Lot # 0000191071
Secondary description: goat anti-mouse secondary antibody
     Dilution: 1/2000
     Diluent Buffer: 3% milk
     Incubation Time: 1 hour
     Incubation Temperature: room temperature
Detection Method:
Detection: ECL (GE, cat # RPN2209, lot # 9838243)
Procedure: Add equal volume of A and B, mix and apply on the membranes for 3-5 min before exposure
Development Time: 40 seconds
Molecular weight of band(s): ~ 48 kDa
Experimental Concerns and Observations: A specific band around 48 kDa was observed
Western Blot GADD153/CHOP NB600-1335
Enlarge
reviewed by:
Wenliang Zhang
WB Rat 09/20/2017
View

Summary

ApplicationWestern Blot
Sample TestedLiver
SpeciesRat
LotA-2
Western Blot GADD153/CHOP NB600-1335
Enlarge
reviewed by:
Fabrice GANKAM
WB Rat 11/02/2014
View

Summary

ApplicationWestern Blot
Sample TestedRat brain
SpeciesRat
CommentsSuprisingly detect 2 bands in rat brain at 16 and around 25 Kda, same pattern seen with antibodies from santacruz and sigma

Blocking

Blocking Detailsmilk 6% in PBST0.02% 2 hrs at RT

Primary Anitbody

Dilution Ratio1/2000 in PBST with milk 3% at 4 degres for 8 hrs

Secondary Antibody

Secondary Descriptiongoat anti mouse HRP rat cross adsorbed
Secondary Manufacturer Cat#Pierce
Secondary Concentration1/20000

Details

Detection NotesECL exposure time of 10 sec using Chemidoc XRS and biorad clarity

Comments

CommentsSuprisingly detect 2 bands in rat brain at 16 and around 25 Kda, same pattern seen with antibodies from santacruz and sigma

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for GADD153/CHOP Antibody (NB600-1335) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB600-1335AF405
Alexa Fluor 488 Labeled NB600-1335AF488
Alexa Fluor 647 Labeled NB600-1335AF647
Alexa Fluor 700 Labeled NB600-1335AF700
Biotin Labeled NB600-1335B
DyLight 350 Labeled NB600-1335UV
DyLight 405 Labeled NB600-1335V
DyLight 488 Labeled NB600-1335G
DyLight 550 Labeled NB600-1335R
DyLight 650 Labeled NB600-1335C
DyLight 680 Labeled NB600-1335FR
DyLight 755 Labeled NB600-1335IR
FITC Labeled NB600-1335F
HRP Labeled NB600-1335H

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Recent Reviews

4.3
5
1
4
2
3
0
2
0
1
0

Anonymous
12/15/2017
Application: WB
Species: Human

Wenliang Zhang
09/20/2017
Application: WB
Species: Rat

Fabrice GANKAM
11/02/2014
Application: WB
Species: Rat

Bioinformatics

Gene Symbol DDIT3
Entrez
Uniprot