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DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31]

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Catalog# ET1703-05

DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31]

Application

  • WB

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

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

种属反应性

Human, Mouse, Rat

验证应用

WB, FC

分子量

Predicted band size: 19 kDa

阳性对照

SW480 cell lysate, HeLa cell lysate, LOVO cell lysate, PC-12 cell lysate, Hela.

偶联

unconjugated

克隆号

JM10-31

RRID

AB_3070363

产品特性

形态

Liquid

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:5,000

  • FC

  • 1:50-1:100

靶点

功能

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.

背景文献

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).

序列相似性

Belongs to the bZIP family.

翻译后修饰

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.

亚细胞定位

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P35638 Human

SwissProt: P35639 Mouse

SwissProt: Q62857 Rat

别名

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

展开

图片

  • Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (<a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</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/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</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 DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (ET1703-05) 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 (ET1703-05) 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/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</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 (ET1703-05) 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.

  • Western blot analysis of DDIT3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</a>, 1/500) was 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 1:200,000 dilution was used for 1 hour at room temperature.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: LOVO cell lysate<br />Lane 2: PC-12 cell lysate<br /><br />Predicted band size: 19 kDa<br />Observed band size: 25 kDa

    Western blot analysis of DDIT3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-05, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: LOVO cell lysate
    Lane 2: PC-12 cell lysate

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

  • Flow cytometric analysis of DDIT3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1703-05" style="font-weight: bold;text-decoration: underline;">ET1703-05</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 DDIT3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-05, 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).

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

引文

Filter:
  1. WB
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  3. IF
  4. IP

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