WB
Western blot analysis of CD31 on different lysates with Mouse anti-CD31 antibody at 1/2,000 dilution. Lane 1: THP-1 cell lysate Lane 2: Jurkat cell lysate Lane 3: U-937 cell lysate Lane 4: HeLa cell lysate (negative) Lane 5: MDA-MB-231 cell lysate (negative) Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 58 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue with Mouse anti-CD31 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of THP-1 (positive) and HeLa (negative) labeling CD31 with Mouse anti-CD31 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-CD31 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.IF-P
mIHC analysis of human lung squamous cell carcinoma tissue (Formalin/PFA-fixed paraffin-embedded sections) with Mouse anti-CD31 antibody at 1/2,000 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.| Product Name | CD31 Mouse Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide (KLH-coupled) within C-terminal residues of human CD31. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG2a |
| Host Species | Mouse |
| Tested Applications | ICC/IFIF-PIHCWB |
| WB:1:500-1:5000 IHC:1:500-1:2000 ICC:1:50-1:200 IF-P:1:1000-1:3000 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PECAM1 |
|---|---|
| Gene Synonyms | CD31 PECA1 GPIIA' PECAM-1 endoCAM CD31/EndoCAM |
| Gene Full Name | platelet and endothelial cell adhesion molecule 1 |
| Gene Summary | The protein encoded by this gene is found on the surface of platelets, monocytes, neutrophils, and some types of T-cells, and makes up a large portion of endothelial cell intercellular junctions. The encoded protein is a member of the immunoglobulin superfamily and is likely involved in leukocyte migration, angiogenesis, and integrin activation. [provided by RefSeq, May 2010] |
| Molecular Weight(MW) | 83kDa(Observed band size: 130kDa) |
| Cellular Localization | Cell junction. Cell membrane. Membrane. |
WB
Western blot analysis of CD31 on different lysates with Mouse anti-CD31 antibody at 1/2,000 dilution. Lane 1: THP-1 cell lysate Lane 2: Jurkat cell lysate Lane 3: U-937 cell lysate Lane 4: HeLa cell lysate (negative) Lane 5: MDA-MB-231 cell lysate (negative) Lysates/proteins at 20 µg/Lane. Exposure time: 1 minute 58 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue with Mouse anti-CD31 antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of THP-1 (positive) and HeLa (negative) labeling CD31 with Mouse anti-CD31 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-CD31 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.
IF-P
mIHC analysis of human lung squamous cell carcinoma tissue (Formalin/PFA-fixed paraffin-embedded sections) with Mouse anti-CD31 antibody at 1/2,000 dilution. The immunostaining was performed with the IRISKit® HyperView mTSA Kit. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.| Application Notes | WB:1:500-1:5000 IHC:1:500-1:2000 ICC:1:50-1:200 IF-P:1:1000-1:3000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM643512