Anti-CD31 antibody [JC/70A]

• 产品名称

  Anti-CD31抗体[JC/70A]
  参阅全部 CD31 一抗

• 描述

  小鼠单克隆抗体[JC/70A] to CD31

• 宿主

  Mouse

• 经测试应用

  适用于: IHC-Fr, WB, IHC-P, Flow Cyt, ICC/IFmore details

• 种属反应性

  与反应: Human
  预测可用于: Cynomolgus monkey

• 免疫原

  Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB: HUVEC and HeLa whole cell lysate. Human spleen and kidney tissue lysate. IHC-P: Human tonsil tissue. ICC/IF: HUVEC cells. Flow Cyt: PBMC.

• 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

• 纯度

  Affinity purified

• 克隆

  单克隆

• 克隆编号

  JC/70A

• 骨髓瘤

  unknown

• 同种型

  IgG1

• 轻链类型

  kappa

The Abpromise guarantee

Abpromise™承诺保证使用ab9498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: 102117792 Cynomolgus monkey

• Entrez Gene: 5175 Human

• Omim: 173445 Human

• SwissProt: P16284 Human

• Unigene: 376675 Human

• Unigene: 514412 Human

• 别名

• CD31 EndoCAM antibody

• EndoCAM antibody

• FLJ34100 antibody

• FLJ58394 antibody

• GPIIA antibody

• GPIIA' antibody

• PECA1 antibody

• PECA1_HUMAN antibody

• Pecam 1 antibody

• PECAM 1 CD31 EndoCAM antibody

• PECAM antibody

• PECAM-1 antibody

• Pecam1 antibody

• Platelet and endothelial cell adhesion molecule 1 antibody

• Platelet endothelial cell adhesion molecule antibody

• Platelet/endothelial cell adhesion molecule 1 antibody

• Adhesion molecule antibody

• CD31 antibody

• CD31 antigen antibody

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [JC/70A] (ab9498)

  IHC image of CD31 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab9498, 0.5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [JC/70A] (ab9498)

  ab9498 staining CD31 in HUVEC cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1%PBS-Tween for 5 minutes 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 ab9498 at 1μg/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 labelled with DAPI (shown in blue).

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

• 

  Flow Cytometry - Anti-CD31 antibody [JC/70A] (ab9498)

  Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab9498 (right) or mouse IgG1κ; (ab170190) isotype (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab9498) or mouse IgG1κ; (ab170190) isotype (1x106 in 100 µL at 0.04 µg/ml) for 30 min on ice.

  The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150117) was used at 1:2000 dilution for 30 min on ice.

  Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on alive cells.

• 

  Western blot - Anti-CD31 antibody [JC/70A] (ab9498)

  All lanes : Anti-CD31 antibody [JC/70A] (ab9498) at 1 µg/ml
  
  Lane 1 : HUVEC whole cell lysate
  Lane 2 : HeLa whole cell lysate
  Lane 3 : Human kidney tissue lysate
  Lane 4 : Human spleen tissue lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Predicted band size: 82 kDa
  Observed band size: 130 kDawhy is the actual band size different from the predicted?
  
  
  
  

  This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab9498 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

• 

  Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [JC/70A] (ab9498)

  IHC image of ab9498 staining in 10% formaldehyde fixed frozen tissue section of human lung.

  Non-specific protein-protein interactions were blocked using TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1 hour at room temperature. The section was then incubated with ab9498 (1μg/ml concentration) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. The section was then incubated with ab150119 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647)) and DAPI for 1 hour at room temperature.

  The DAPI only control (no antibody) inset shows no autofluorescence, demonstrating that any Alexa Fluor® 647 signal is derived directly from bound ab9498. 

  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [JC/70A] (ab9498)

  IHC image of CD31 staining in human spleen formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9498, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

   

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Flow Cytometry - Anti-CD31 antibody [JC/70A] (ab9498)

  Overlay histogram showing Jurkat cells stained with ab9498 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9498, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min) used under the same conditions.
  
  Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [JC/70A] (ab9498)

  ab9498 stained in HUVEC cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab9498 at 5 µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150080 (pseudo-colored red) and ab150117 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [JC/70A] (ab9498)

  ab9498 stained HUVEC cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9498 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody [JC/70A] (ab9498)This image is courtesy of an anonymous Abreview

  ab9498 at 1/100 staining human lymph node tissue sections by IHC-P. The tissue was paraformaldehyde fixed and a heat based antigen retrieval step was performed. The tissue was then blocked with serum and incubated with ab9498 overnight. An HRP conjugated goat antibody was used as the secondary.

  See Abreview