Anti-DDIT3 antibody [9C8]

• 产品名称

  Anti-CHOP抗体[9C8]
  参阅全部 CHOP 一抗

• 描述

  小鼠单克隆抗体[9C8] to CHOP

• 宿主

  Mouse

• 经测试应用

  适用于: WB, ICC/IFmore details

• 种属反应性

  与反应: Mouse, Human
  预测可用于: Rat

• 免疫原

  Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

• 表位

  ab11419 has been shown to recognize an epitope in the N-terminal region of DDIT3.

• 阳性对照

• WB: SW480 cell lysates, HeLa cells treated with 2ug/ml tunicamycin for 4 hours, NIH3T3 cell lysate, Wild-type HeLa Treated Tunicamycin cell lysate ICC/IF: HeLa (untreated and tunicamycin-treated).

• 常规说明

  
  

  This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

• 存储溶液

  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine

• 浓度

• 10 µg 浓度为 1 mg/ml

• 100 µg 浓度为 1 mg/ml

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  9C8

• 同种型

  IgG2b

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab11419于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

数据库链接

• Entrez Gene: 1649 Human

• Entrez Gene: 13198 Mouse

• Entrez Gene: 29467 Rat

• Omim: 126337 Human

• SwissProt: P35638 Human

• SwissProt: P35639 Mouse

• SwissProt: Q62857 Rat

• Unigene: 505777 Human

• Unigene: 728989 Human

• Unigene: 110220 Mouse

• Unigene: 11183 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

• CHOP10 antibody

• DDIT 3 antibody

• DDIT-3 antibody

• Ddit3 antibody

• DDIT3_HUMAN antibody

• DNA Damage Inducible Transcript 3 antibody

• DNA damage-inducible transcript 3 protein antibody

• GADD 153 antibody

• GADD153 antibody

• Growth Arrest and DNA Damage Inducible Protein 153 antibody

• Growth arrest and DNA damage inducible protein GADD153 antibody

• Growth arrest and DNA damage-inducible protein GADD153 antibody

• MGC4154 antibody

• 

  Immunocytochemistry - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous Abreview

  Immunocytochemistry analysis of formaldehyde-fixed HT29 cells permeabilized with 0.1% TritonX-100 in PBS for 10min staining with ab11419 at 1/500. Secondary antibody was ImmPRESS® HRP Goat Anti-Mouse IgG Polymer Detection. Samples were incubated with the primary antibody with Immunofluorescence Antibody Dilution Buffer for 18 hours at 4°C. Blocking was done using Peroxidase Blocking Solution, BLOXALL for 20 minutes at 20°C.

  See Abreview

• 

  Western blot - Anti-DDIT3 antibody [9C8] (ab11419)

  All lanes : Anti-CHOP antibody [9C8] (ab11419)
  
  Lane 1 : Wild-type HeLa Vehicle Control Tunicamycin cell lysate
  Lane 2 : Wild-type HeLa Treated Tunicamycin cell lysate
  Lane 3 : DDIT3 knockout HeLa Vehicle Control Tunicamycin cell lysate
  Lane 4 : DDIT3 knockout HeLa Treated Tunicamycin cell lysate
  
  Lysates/proteins at 20 µg/ml per lane.
  
  Performed under reducing conditions.
  
  Predicted band size: 19 kDa
  Observed band size: 25 kDawhy is the actual band size different from the predicted?
  
  
  
  

  False colour image of Western blot: Anti-DDIT3 antibody [9C8] staining at 5µg/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab11419 was shown to bind specifically to DDIT3. A band was observed at 25 kDa in wild-type y cell lysates with no signal observed at this size in DDIT3 knockout cell line ab265760 (knockout cell lysate ab256889). To generate this image, wild-type and DDIT3 knockout y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

• 

  Western blot - Anti-DDIT3 antibody [9C8] (ab11419)

  All lanes : Anti-CHOP antibody [9C8] (ab11419) at 5 µg/ml
  
  Lane 1 : Wild-type SW480 cell lysate
  Lane 2 : DDIT3 knockout SW480 cell lysate
  Lane 3 : Untreated HeLa cell lysate
  Lane 4 : HeLa + DMSO control cell lysate
  Lane 5 : HeLa + tunicamycin (20ug/mL,4 hours) cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Predicted band size: 19 kDa
  Observed band size: 26 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Lanes 1 - 5: Merged signal (red and green). Green - ab11419 observed at 26 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

  ab11419 was shown to react with DDIT3 in wild-type SW480 cells in western blot with loss of signal observed in DDIT3 knockout sample. Wild-type and DDIT3 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab11419 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)

  ab11419 staining DDIT3 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Tunicamycin 1.5μM, 6 hours (ab120296). 

  The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% Triton-X for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab11419 at 5μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Western blot - Anti-DDIT3 antibody [9C8] (ab11419)

  All lanes : Anti-CHOP antibody [9C8] (ab11419) at 5 µg/ml
  
  Lane 1 : HeLa w/c control cell lysate at 40 µg
  Lane 2 : HeLa cells treated with 2ug/ml tunicamycin for 4 hours, whole cell lysate cell lysate at 40 µg
  Lane 3 : HeLa cells treated with 20ug/ml tunicamycin for 4 hours, whole cell lysate cell lysate at 40 µg
  Lane 4 : HepG2 cell lysate at 20 µg
  Lane 5 : NIH3T3 cell lysate at 20 µg
  
  Performed under reducing conditions.
  
  Predicted band size: 19 kDa
  
  
  
  

  Lanes 1 - 5: Merged signal (red and green). Green - ab11419 observed at 27 kDa. Red - loading control, Rabbit anti Actin observed at 42kDa.

  ab11419 was shown to react with DDIT3 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab11419 and Rabbit anti Actin overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

• 

  Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous Abreview

  All lanes : Anti-CHOP antibody [9C8] (ab11419) at 1/1000 dilution
  
  All lanes : Mouse hepatocyte whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Secondary
  All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 19 kDa
  Observed band size: 27 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 5 minutes
  
  
  

  Treated with 20µg/ml poly(I:C).

  See Abreview

• 

  Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous Abreview

  All lanes : Anti-CHOP antibody [9C8] (ab11419) at 1/500 dilution (in TBST + 2.5% milk for 16 hours at 4°C)
  
  Lane 1 : Whole cell lystate of Mouse 3T3 cells
  Lane 2 : Whole cell lystate of Mouse 3T3 cells treated with tunicamycin for 24 hours
  
  Lysates/proteins at 50 µg per lane.
  
  Secondary
  All lanes : An HRP-conjugated Goat anti-mouse IgG monoclonal at 1/2000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 19 kDa
  Observed band size: 31 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 2 minutes
  
  
  Blocking Step: 5% Milk for 2 hours at 22°C

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)

  ab11419 staining DDIT3 in SK-N-SH (human neuroblastoma cell line) cells treated with deltamethrin (ab141019), by ICC/IF. Increase of DDIT3 expression correlates with increased concentration of deltamethrin, as described in literature.
  The cells were incubated at 37°C for 48 hours in media containing different concentrations of ab141019 (deltamethrin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab11419 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight® 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.