Phospho-DDIT3/CHOP (Ser30) Antibody - #AF3277

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Product: Phospho-DDIT3/CHOP (Ser30) Antibody
Catalog: AF3277
Description: Rabbit polyclonal antibody to Phospho-DDIT3/CHOP (Ser30)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.: 19~30kD; 19kD(Calculated).
Uniprot: P35638
RRID: AB_2834505

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Related Downloads
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(89%), Sheep(100%), Rabbit(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
Phospho-DDIT3 (Ser30) Antibody detects endogenous levels of DDIT3 only when phosphorylated at Serine 30.
RRID:
AB_2834505
Cite Format: Affinity Biosciences Cat# AF3277, RRID:AB_2834505.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

C/EBP homologous protein; C/EBP Homology Protein; C/EBP zeta; C/EBP-homologous protein 10; C/EBP-homologous protein; CCAAT/enhancer binding protein homologous protein; CEBPZ; CHOP 10; CHOP; CHOP-10; CHOP10; DDIT 3; DDIT-3; Ddit3; DDIT3_HUMAN; DNA Damage Inducible Transcript 3; DNA damage-inducible transcript 3 protein; GADD 153; GADD153; Growth Arrest and DNA Damage Inducible Protein 153; Growth arrest and DNA damage inducible protein GADD153; Growth arrest and DNA damage-inducible protein GADD153; MGC4154;

Immunogens

Immunogen:
Uniprot:

P35638(DDIT3_HUMAN) >>Visit HPA database.

Gene(ID):
DDIT3(1649)
Description:
CHOP a transcriptional-regulatory protein of the bZIP family. Inhibits the DNA-binding activity of C/EBP and LAP by forming heterodimers that cannot bind DNA. May play an important role in melanoma progression. CK2-mediated phosphorylation inhibits its transcriptional activity.
Sequence:
MAAESLPFSFGTLSSWELEAWYEDLQEVLSSDENGGTYVSPPGNEEEESKIFTTLDPASLAWLTEEEPEPAEVTSTSQSPHSPDSSQSSLAQEEEEEDQGRTRKRKQSGHSPARAGKQRMKEKEQENERKVAQLAEENERLKQEIERLTREVEATRRALIDRMVNLHQA

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Horse
89
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P35638 As Substrate

Site PTM Type Enzyme
S14 Phosphorylation P68400 (CSNK2A1)
S15 Phosphorylation P68400 (CSNK2A1)
S30 Phosphorylation P68400 (CSNK2A1)
S31 Phosphorylation P68400 (CSNK2A1)
S49 Phosphorylation
T54 Phosphorylation
T64 Phosphorylation
S79 Phosphorylation Q16539 (MAPK14)
S82 Phosphorylation Q16539 (MAPK14)

Research Backgrounds

Function:

Multifunctional transcription factor in ER stress response. Plays an essential role in the response to a wide variety of cell stresses and induces cell cycle arrest and apoptosis in response to ER stress. Plays a dual role both as an inhibitor of CCAAT/enhancer-binding protein (C/EBP) function and as an activator of other genes. Acts as a dominant-negative regulator of C/EBP-induced transcription: dimerizes with members of the C/EBP family, impairs their association with C/EBP binding sites in the promoter regions, and inhibits the expression of C/EBP regulated genes. Positively regulates the transcription of TRIB3, IL6, IL8, IL23, TNFRSF10B/DR5, PPP1R15A/GADD34, BBC3/PUMA, BCL2L11/BIM and ERO1L. Negatively regulates; expression of BCL2 and MYOD1, ATF4-dependent transcriptional activation of asparagine synthetase (ASNS), CEBPA-dependent transcriptional activation of hepcidin (HAMP) and CEBPB-mediated expression of peroxisome proliferator-activated receptor gamma (PPARG). Inhibits the canonical Wnt signaling pathway by binding to TCF7L2/TCF4, impairing its DNA-binding properties and repressing its transcriptional activity. Plays a regulatory role in the inflammatory response through the induction of caspase-11 (CASP4/CASP11) which induces the activation of caspase-1 (CASP1) and both these caspases increase the activation of pro-IL1B to mature IL1B which is involved in the inflammatory response.

PTMs:

Ubiquitinated, leading to its degradation by the proteasome.

Phosphorylation at serine residues by MAPK14 enhances its transcriptional activation activity while phosphorylation at serine residues by CK2 inhibits its transcriptional activation activity.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Present in the cytoplasm under non-stressed conditions and ER stress leads to its nuclear accumulation.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

Heterodimer. Interacts with TCF7L2/TCF4, EP300/P300, HDAC1, HDAC5 and HDAC6. Interacts with TRIB3 which blocks its association with EP300/P300. Interacts with FOXO3, CEBPB and ATF4. Interacts with isoform AltDDIT3 of DDIT3.

Family&Domains:

The N-terminal region is necessary for its proteasomal degradation, transcriptional activity and interaction with EP300/P300.

Belongs to the bZIP family.

Research Fields

· Cellular Processes > Cell growth and death > Apoptosis.   (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Genetic Information Processing > Folding, sorting and degradation > Protein processing in endoplasmic reticulum.   (View pathway)

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

References

1). Aberrant activation of p53-TRIB3 axis contributes to diabetic myocardial insulin resistance and sulforaphane protection. Journal of advanced research, 2024 (PubMed: 39069209) [IF=10.7]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 7. p53-induced TRIB3 upregulation depends on the transcription factor CHOP. (A) The mRNA level of Trib3 was analyzed by qRT-PCR in H9c2 cells (n = 3). (B) Transcription factors were predicted to bind to Trib3 by FIMO. Predictive intersection of rat and mouse. Screening condition q-value < 0.05. (C) CHOP binding motif provided by the JASPAR database. Cell-based luciferase reporter assays were detected in H9c2 cells co-transfected with luciferase-expressing vectors driven by the Trib3 promoter or the mutant promoters, together with an empty plasmid as the control for 24 h (n = 3). (D, E) ChIP analysis was performed with anti-CHOP antibody or IgG in H9c2 cells. The precipitated chromatin was analyzed by qRT-PCR (n = 3). (F) Proteins interacting between p53 and CHOP were retrieved from the STRING database ( https://string-db.org/ ). (G) Interaction of p53 and CHOP in H9c2 cells determined by co-IP. (H) Co-localization of p53 and CHOP was determined by immunofluorescent staining in H9c2 cells. (I) The expression of p-CHOP and CHOP in cardiac tissues was examined by western blot (n = 6). (J) Interaction of CHOP and AMPKα in H9c2 cells demonstrated by co-IP. (K, L) The p-CHOP and CHOP level was detected by western blot analysis after transfection with NC-shRNA or Ampk-shRNA in H9c2 cells (n = 3). (M) CHOP was immunoprecipitated from H9c2 cells of different groups and measured using western blot analysis with an anti-ubiquitin antibody. (N) The immunoblotting of immunoprecipitated CHOP with antibody-recognized ubiquitin of different groups in H9c2 cells. (O) ChIP analysis was performed with anti-CHOP antibody or IgG in H9c2 cells. The precipitated chromatin was detected by qRT-PCR (n = 3). (P) Diagram showing the experimental design for the mice study. (Q) Representative M-mode echocardiographic images. (R) Western blot analysis of p-AKT, p-GSK-3β, and p-GS in cardiac tissues (n = 6). Data shown in the graphs represents the means ± SD. *P < 0.05; N.S., not significant.

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