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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for CD31/PECAM-1 Antibody - BSA Free

Immunogen

This CD31/PECAM-1 Antibody was developed against a genomic peptide made to an internal region of human CD31/PECAM1 (within residues 100-300) [Swiss-Prot P16284].

Localization

Membrane; Single-pass type I membrane protein. Cell junction. Note: Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

82.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CD31/PECAM-1 Antibody - BSA Free

Immunocytochemistry/Immunofluorescence: CD31/PECAM-1 Antibody [NBP1-71663] - Staining of mouse glioma tissue using CD31/PECAM-1 Antibody (NBP1-71663) at a dilution of 1:2500. Image provided by Dr. Eric Woolf of Dignity Health.
Simple Western: CD31/PECAM-1 Antibody [NBP1-71663] - Image shows a specific band for CD31/PECAM 1 in 0.5 mg/mL of HUVEC lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Immunohistochemistry: CD31/PECAM-1 Antibody [NBP1-71663] - 5-HTR1A agonist enhances neovascularization during skin wound healing (excisional punch biopsy model). CD31/PECAM-1 endothelial cell immunohistochemical staining at day 5 (A,B) and day 7 (E,F), image scale bars represented at 50 um. Results were presented as mean +/- SEM (n = 5 images per wound). Two-way ANOVA was performed and significance levels were set at *P < 0.05, **P < 0.01. Image collected and cropped by CiteAb from the following publication (https://www.frontiersin.org/article/10.3389/fphar.2018.01406/full), licensed under a CC-BY license.

Applications for CD31/PECAM-1 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

1:1000

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:50

Immunohistochemistry-Frozen

reported in scientific literature (PMID 35879332)

Immunohistochemistry-Paraffin

1:50

Simple Western

1:1000

Western Blot

1:10000
Application Notes
In Western Blot a band is seen ~150 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. Customer feedback has been negative on cryosections.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using NBP1-71663 in the following applications:

Published Applications

Read 16 publications using NBP1-71663 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS, 30% Glycerol

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD31/PECAM-1

Cluster of differentiation (CD31), Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1), is a 30kDa type I transmembrane glycoprotein expressed by endothelial cells, platelets, leukocytes and some lymphocytes (e.g., B and T-cell subsets). Structurally, PECAM-1 consists of six extracellular immunoglobulin-like domains which serve to support various functions including homophilic binding, maintaining high expression of PECAM-1 within endothelial intercellular junctions, and supporting endothelial cell migration (1,2). PECAM's homophilic binding underscores its role in vascular barrier integrity and leukocyte trans-endothelial movement. However, PECAM may also engage in heterophilic interactions with molecules such as glycosaminoglycans (GAG), integrin alpha 5 beta 3, CD38 in lymphocytes and CD177/PR3 in neutrophils (2).

PECAM's intracellular cytoplasmic domain consists of a sequence of 118 amino acids and contains serine and tyrosine (also referred to as immunoreceptor tyrosine-based inhibitory motifs-ITIMs) residues, which may be phosphorylated upon cellular stimulation (3). ITIMs are phosphorylated by Src-family kinases and non-Src family kinases (e.g., Csk), leading to a conformational change which supports interactions with Src homology 2 (SH2) domain containing proteins such as protein-tyrosine phosphatase, SHP-2 (1,2). Formation of SHP-2/PECAM-1 complexes induces endothelial cell migration through the dephosphorylation of focal adhesion kinase and regulation of RhoA activity (1). Signaling downstream of ITIM tyrosine phosphorylations also plays a role in PECAM's anti-apoptotic activity, a function which is independent of its interaction with SHP-2. In platelets and leukocytes, phosphorylation of PECAM's cytosolic domain is inhibitory, preventing their activation.

References

1. Lertkiatmongkol, P., Liao, D., Mei, H., Hu, Y., & Newman, P. J. (2016). Endothelial functions of PECAM-1 (CD31). Current Opinion in Hematology. https://doi.org/10.1097/MOH.0000000000000239.Endothelial

2. Privratsky, J. R., & Newman, P. J. (2014). PECAM-1: Regulator of endothelial junctional integrity. Cell and Tissue Research. https://doi.org/10.1007/s00441-013-1779-3

3. Newman, P. J., & Newman, D. K. (2003). Signal transduction pathways mediated by PECAM-1: New roles for an old molecule in platelet and vascular cell biology. Arteriosclerosis, Thrombosis, and Vascular Biology. https://doi.org/10.1161/01.ATV.0000071347.69358.D9

Long Name

Platelet Endothelial Cell Adhesion Molecule 1

Alternate Names

CD31, EndoCAM, PECA1, PECAM-1, PECAM1

Entrez Gene IDs

5175 (Human); 18316 (Mouse)

Gene Symbol

PECAM1

UniProt

Product Documents for CD31/PECAM-1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for CD31/PECAM-1 Antibody - BSA Free

Manufactured by Genomic Antibody Technology™. GAT FAQs

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

View specific protocols for CD31/PECAM-1 Antibody - BSA Free (NBP1-71663):

[[URL:https://www.novusbio.com/products/cd31-pecam-1-antibody_nbp1-71663]][[C… Antibody]]
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.
[[URL:https://www.novusbio.com/products/cd31-pecam-1-antibody_nbp1-71663]][[C… antibody]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

FAQs for CD31/PECAM-1 Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: I'm interested in purchasing primary CD31/PECAM1 antibody for immunohistochemistry staining on formalin fixed paraffin embedded mouse samples. You have two antibodies that would work for paraffin sections, NBP1-71663 and NB100-92205, and both are rabbit polyclonal antibody. Which one would you recommend I purchase?

    A: I would recommend NBP1-71663 as the image is better than NB100-92205.

  • Q: I am looking for references concerning the use of NBP2-15797, NBP1-47913 and NB100-2284 in IHC on PFA- fixed section.

    A: Among your mentioned catalog numbers (NBP2-15797, NBP1-47913 and NB100-2284), we have published references for NB100-2284 only. Hylander et al. 2013 (Journal of Translational Medicine 2013, 11:110) have cited the IHC-P use of NB100-2284 and this paper has also explained a nice detailed IHC-P protocol that you may follow if you want to. NBP1-71663 is our bestselling catalog number among all CD31/PECAM1 antibodies and it has been cited in two publications, both explaining its use in IHC-P assays (Pratheeshkumar et al. 2012 PLoS One. 2012;7(10):e47516; and Pratheeshkumar et al. 2012 PLoS One. 2012;7(12):e52279)

Showing  1 - 2 of 2 FAQs Showing All
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