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CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750369

CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31] - BSA and Azide free

Application

  • WB

  • FC(Intra)

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human DDIT3 aa 135-169 / 169.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, FC(Intra)

Target Molecular Weight

Predicted band size: 19 kDa

Positive Control

SW480 cell lysate, HeLa cell lysate, Hela.

Conjugation

unconjugated

Clone Number

JM10-31

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • FC(Intra)

  • 1:500

Target

Function

DNA damage-inducible transcript 3, also known as C/EBP homologous protein (CHOP), is a pro-apoptotic transcription factor that is encoded by the DDIT3 gene. It is a member of the CCAAT/enhancer-binding protein (C/EBP) family of DNA-binding transcription factors. The protein functions as a dominant-negative inhibitor by forming heterodimers with other C/EBP members, preventing their DNA binding activity. The protein is implicated in adipogenesis and erythropoiesis and has an important role in the cell's stress response. The regulation of CHOP expression plays an important role in metabolic diseases and in some cancers through its function in mediating apoptosis. The regulation of CHOP expression could be a potential approach to affecting cancer cells through the induction of apoptosis. In the intestinal epithelium, CHOP has been demonstrated to be downregulated under inflammatory conditions (in inflammatory bowel diseases and experimental models of colitis). In this context, CHOP seems to rather regulate the cell cycle than apoptotic processes. Mutations or fusions of CHOP (e.g. with FUS to form FUS-CHOP) can cause Myxoid liposarcoma.

Background References

1. Sun XY et al. Valproate attenuates diabetic nephropathy through inhibition of endoplasmic reticulum stress-induced apoptosis. Mol Med Rep 13:661-8 (2016).

2. Greenwood M et al. Transcription Factor CREB3L1 Regulates Endoplasmic Reticulum Stress Response Genes in the Osmotically Challenged Rat Hypothalamus. PLoS One 10:e0124956 (2015).

Sequence Similarity

Belongs to the bZIP family.

Post-translational Modification

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.

UNIPROT

SwissProt: P35638 Human

SwissProt: P35639 Mouse

SwissProt: Q62857 Rat

Synonyms

C/EBP homologous protein antibody

C/EBP Homology Protein antibody

C/EBP zeta antibody

C/EBP-homologous protein 10 antibody

C/EBP-homologous protein antibody

CCAAT/enhancer binding protein homologous protein antibody

CEBPZ antibody

CHOP 10 antibody

CHOP antibody

CHOP-10 antibody

Expand

Images

  • Western blot analysis of CHOP/DDIT3 on different lysates with Rabbit anti-CHOP/DDIT3 antibody (<a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>) at 1/5,000 dilution.<br /><br />Lane 1: SW480 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br /><br />Predicted band size: 19 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of CHOP/DDIT3 on different lysates with Rabbit anti-CHOP/DDIT3 antibody (HA750369) at 1/5,000 dilution.

    Lane 1: SW480 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)

    Predicted band size: 19 kDa
    Observed band size: 27 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750369) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (<a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>) at 1/1,000 dilution.<br /><br />Lane 1: SW480-si NT cell lysate<br />Lane 2: SW480-si DDIT3 cell lysate<br /><br />Lysates/proteins at 3 µg/Lane.<br /><br />Predicted band size: 19 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</a> was shown to specifically react with DDIT3 in SW480-si NT cells. Weakened band was observed when SW480-si DDIT3 sample was tested. SW480-si NT and SW480-si DDIT3 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</a>, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (HA750369) at 1/1,000 dilution.

    Lane 1: SW480-si NT cell lysate
    Lane 2: SW480-si DDIT3 cell lysate

    Lysates/proteins at 3 µg/Lane.

    Predicted band size: 19 kDa
    Observed band size: 25 kDa

    Exposure time: 5 minutes;

    4-20% SDS-PAGE gel.

    ET1703-05 was shown to specifically react with DDIT3 in SW480-si NT cells. Weakened band was observed when SW480-si DDIT3 sample was tested. SW480-si NT and SW480-si DDIT3 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1703-05, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of CHOP/DDIT3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of CHOP/DDIT3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (HA750369, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Western blot analysis of CHOP/DDIT3 on different lysates with Rabbit anti-CHOP/DDIT3 antibody (<a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>) at 1/5,000 dilution.<br /><br />Lane 1: C2C12 cell lysate<br />Lane 2: C2C12 treatment with 300 nM thapsigargin for 18h cell lysate<br />Lane 3: C6 cell lysate<br />Lane 4: C6 treatment with 300 nM thapsigargin for 18h cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 20 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA750369" style="font-weight: bold;text-decoration: underline;">HA750369</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 19 kDa<br />Observed band size: 27 kDa

    Western blot analysis of CHOP/DDIT3 on different lysates with Rabbit anti-CHOP/DDIT3 antibody (HA750369) at 1/5,000 dilution.

    Lane 1: C2C12 cell lysate
    Lane 2: C2C12 treatment with 300 nM thapsigargin for 18h cell lysate
    Lane 3: C6 cell lysate
    Lane 4: C6 treatment with 300 nM thapsigargin for 18h cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA750369, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 19 kDa
    Observed band size: 27 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [PSH07-65]

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CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31]

Application: WB,FC(Intra)

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated

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CHOP/DDIT3 Recombinant Rabbit Monoclonal Antibody [PSH07-65] - BSA and Azide free

Application: WB,IF-Cell,FC(Intra)

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Conjugate: unconjugated