GADD153/CHOP Antibody (9C8) - BSA Free

Novus Biologicals | Catalog # NB600-1335

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Primate

Cited:

Human, Mouse, Rat, Primate

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Gel Super Shift Assays, Knockdown Validated

Cited:

Immunohistochemistry-Paraffin, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chemotaxis, Gel Supershift Assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Kappa Clone # 9C8

Format

BSA Free
View all GADD153 Primary Antibodies »

Product Specifications

Immunogen

Full length mouse CHOP/GADD153 [Swiss-Prot# P35639]

Epitope

The N-terminal region of CHOP/GADD153.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID:32828953). Human, mouse, rat and primate.

Localization

Nuclear

Marker

ER Stress Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Kappa

Theoretical MW

19 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.

Scientific Data Images for GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

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]

Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335]

Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335] - FFPE tissue section of mouse brain using 1:100 dilution of GADD153/CHOP antibody. The signal was developed using HRP-DAB based detection method which followed counterstaining of the nuclei with hematoxylin. The antibody generated a cytoplasmic and nuclear staining of CHOP in various cell types in the tested section.
Flow Cytometry: GADD153/CHOP Antibody (9C8) [NB600-1335]

Flow Cytometry: GADD153/CHOP Antibody (9C8) [NB600-1335]

Flow Cytometry: GADD153/CHOP Antibody (9C8) [NB600-1335] - An intracellular stain was performed on SK-MEL-28 cells with GADD153/CHOP Antibody (9C8) NB600-1335 (blue) and a matched isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).
Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

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).
Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

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 with tunicamycin for 10 hours.
Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

Western Blot: GADD153/CHOP Antibody (9C8) [NB600-1335]

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]

Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335]

Immunohistochemistry-Paraffin: GADD153/CHOP Antibody (9C8) [NB600-1335] - FFPE tissue section of mouse brain using 1:100 dilution of GADD153/CHOP antibody. The signal was developed using HRP-DAB based detection method which followed counterstaining of the nuclei with hematoxylin. The antibody generated a cytoplasmic and nuclear staining of CHOP in various cell types in the tested section.
Simple Western: GADD153/CHOP Antibody (9C8) [NB600-1335]

Simple Western: GADD153/CHOP Antibody (9C8) [NB600-1335]

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 separation system.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

The TUG1‐miR‐144‐3p‐Nrf2 axis regulates H/R‐induced cell apoptosis by adjusting oxidative stress and endoplasmic reticulum stress in vitro. A, B, TUNEL stain and quantitative analysis for H/R‐injured TCMK cells with different transfections (n = 3). C–J, The expression levels of Nrf2, H0‐1, CHOP, GRP78, Bax, Bcl‐2 and cleaved‐caspase3 in H/R‐injured TCMK cells with different transfections were measured by Western blot analysis. Representative protein bands are shown in F, and the quantitative analysis of protein expression is shown in C‐E, G‐J (n = 3). K, L, The levels of SOD and MDA in different transfection groups (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

MiR‐144‐3p regulates H/R‐induced Nrf2‐HO‐1 signaling pathway activation, oxidative stress, mitochondria and endoplasmic reticulum functions in vitro. A–C, The expression levels of Nrf2 and HO‐1 in TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment. Representative bands are shown in A, and the quantitative analysis of protein expression level is shown in B, C (n = 3). D, E, The SOD and MDA levels in cellular supernatant of TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment (n = 3). F–I, The expression levels of CHOP, GRP78 and Cytochrome C in TCMK cells transfected with miR‐144‐3p mimics or inhibitor after H/R treatment. Representative bands are shown in F, and the quantitative analysis of protein expression level is shown in G–I (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

UPR Western blotting in vitro (A). (B) ASC-CO and ASC-CM reduced the expression of BIP under exposure to CoCl2, incubations with tunicamycin, and CoCl2 were used as controls (C) ASC-CO and ASC-CM reduced the protein expression of ATF6 under CoCl2, incubations with tunicamycin, and CoCl2 were used as controls (D) ASC-CO 57 and ASC-CM reduced the protein expression of XBP1, incubations with tunicamycin, and CoCl2. (E) The protein expression of CHOP and tunicamycin remained unchanged with ASC-CO and ASC-CM as compared to controls. (n = 3, ANOVA ns, not significant; * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/24/24/17197), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

Knocking down Nrf2 reversed the miR‐144‐3p inhibitor‐alleviated H/R‐induced apoptosis, oxidative stress, mitochondrial damage and endoplasmic reticulum stress in vitro. A–c, F–K, The protein expression level of Nrf2, HO‐1, cleaved‐caspase3, Bax, Bcl‐2, CHOP, GRP78 and Cytochrome C in H/R‐injured TCMK cells transfected with miR‐144‐3p inhibitor or si‐Nrf2. Representative bands are shown in A, and the quantitative analysis of protein expression is shown in B, C, F–K (n = 3). D, E, The SOD and MDA levels in cellular supernatants in H/R‐injured TCMK cells transfected with miR‐144‐3p inhibitor or si‐Nrf2 (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01, NS, no significant difference Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

TUG1 plays an important role in H/R‐induced cell apoptosis possibly through regulating the Nrf2‐HO‐1 pathway, oxidative stress, mitochondrial damage and endoplasmic reticulum stress via targeting miR‐144‐3p. A, The expression of miR‐144‐3p in TUG1‐overexpressing or TUG1‐knockdown TCMK cells detected by RT‐qPCR (n = 3). B, C, The levels of SOD and MDA in cellular supernatant (n = 3). D–J, The relative expression levels of Nrf2, HO‐1, GRP78, CHOP, Cyt C and cleaved‐caspase3 were examined. Representative protein bands are shown in D, and the quantitative analysis of protein expression is shown in E‐J (n = 3). All data are expressed as the mean +/- SD; data comparisons between multiple groups were performed using one‐way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34547172), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

ASCs lead to changes in UPR expression in vivo (A) The comparison of the left and right legs—with DLFA- of the two groups of mice are shown (B) Ratio between BIP expression between left and right leg with and without ASCs (C) Ratio of ATF6 expression between left and right leg with and without ASCs (D) Ratio of CHOP expression between left and right leg with and without ASCs (E) Ratio of XBP1 expression between left and right leg with and without ASCs (n = 3, ANOVA, * p ≤ 0.05, ** p ≤ 0.01). Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/1422-0067/24/24/17197), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

Response to ER Stress Mediates the Reprogramming Facilitation by Surf4. (A) The RNA level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. Relative expression of these genes relative to beta ‐actin (n = 3, average +/-SEM). (B) The protein level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. (C) Kinetics of Oct4‐GFP+colony formation with or without exogenous Surf4 and sXbp1‐ delta DBD during reprogramming. (D) The number of Oct4‐GFP+ colonies and the percentage of Oct4‐GFP+ cells induced by OSKM plus Surf4 and sXbp1‐ delta DBD. (E) Morphology of the primary colonies induced by OSKM plus Surf4 and sXbp1‐ delta DBD. Scale bars, 1000 μm. Magnification: ×40. (F) AP staining of the primary iPS colonies. See also Figure S4 and Table S1 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34585448), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
GADD153/CHOP Antibody (9C8) - BSA Free

Western Blot: GADD153/CHOP Antibody (9C8) - BSA Free [NB600-1335] -

Response to ER Stress Mediates the Reprogramming Facilitation by Surf4. (A) The RNA level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. Relative expression of these genes relative to beta ‐actin (n = 3, average +/-SEM). (B) The protein level of ER stress‐related genes on day 3 of reprogramming with or without exogenous Surf4. (C) Kinetics of Oct4‐GFP+colony formation with or without exogenous Surf4 and sXbp1‐ delta DBD during reprogramming. (D) The number of Oct4‐GFP+ colonies and the percentage of Oct4‐GFP+ cells induced by OSKM plus Surf4 and sXbp1‐ delta DBD. (E) Morphology of the primary colonies induced by OSKM plus Surf4 and sXbp1‐ delta DBD. Scale bars, 1000 μm. Magnification: ×40. (F) AP staining of the primary iPS colonies. See also Figure S4 and Table S1 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34585448), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for GADD153/CHOP Antibody (9C8) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1:10 - 1:500

Simple Western

1:250
Application Notes
Use in Knockdown Validated reported in scientific literature (PMID:32828953)In Western blot a band can be seen at approx. 29 Knockdown Validateda. 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.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 1.0 mg/mL, separated by Size, antibody dilution of 1:250, apparent MW was 34 kDa. 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. Use in chromatin immunoprecipitation reported in scientific literature (PMID: 30962207). Use in ELISA reported in scientific literature (PMID: 29915575). Use in FLOW reported in scientific literature (PMID: 8650547).

Reviewed Applications

Read 3 reviews rated 4.3 using NB600-1335 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: GADD153

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.

Long Name

Growth Arrest and DNA Damage-inducible Protein GADD153

Alternate Names

CEBP zeta, CHOP, CHOP10, DDIT3

Entrez Gene IDs

1649 (Human); 13198 (Mouse)

Gene Symbol

DDIT3

UniProt

P35638, P35639 (Mouse)

Additional GADD153 Products

Product Documents for GADD153/CHOP Antibody (9C8) - BSA Free

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Product Specific Notices for GADD153/CHOP Antibody (9C8) - BSA Free

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.

Citations for GADD153/CHOP Antibody (9C8) - BSA Free

Customer Reviews for GADD153/CHOP Antibody (9C8) - BSA Free (3)

4.3 out of 5
3 Customer Ratings
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Showing  1 - 3 of 3 reviews Showing All
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  • GADD153/CHOP Antibody (9C8)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: human liver and primary hepatocytes
    Species: Human
    Verified Customer | Posted 12/15/2017
    Review for anti-CHOP antibody (NB600-1335)
    Name: 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
    GADD153/CHOP Antibody (9C8) - BSA Free NB600-1335
  • GADD153/CHOP Antibody (9C8)
    Name: Wenliang Zhang
    Application: Western Blot
    Sample Tested: Liver
    Species: Rat
    Verified Customer | Posted 09/20/2017
    Ethanol feeding increases CHOP expression.
    GADD153/CHOP Antibody (9C8) - BSA Free NB600-1335
  • Name: Fabrice GANKAM
    Application: Western Blot
    Sample Tested: Rat brain
    Species: Rat
    Verified Customer | Posted 11/02/2014
    GADD153/CHOP Antibody (9C8) - BSA Free NB600-1335

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