CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-80641

Clone JC/70A was used by HLDA to establish CD designation.
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Key Product Details

Species Reactivity

Human, Mouse, Feline, Rabbit

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Flow (Cell Surface), Dual RNAscope ISH-IHC, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # JC/70A

Format

Azide and BSA Free
View all CD31/PECAM-1 Primary Antibodies »

Product Specifications

Immunogen

This CD31/PECAM-1 Antibody (JC/70A) was developed against a membrane preparation of a spleen from a patient with hairy cell leukemia.

Reactivity Notes

Rabbit (PMID: 21533193) and Mouse (PMID: 29700126) reactivity reported in scientific literature.

Localization

Cell membrane.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

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 (JC/70A) - Azide and BSA Free

Western Blot Detection of CD31/PECAM-1 in Multiple Cell Lysates

Western Blot Detection of CD31/PECAM-1 in Multiple Cell Lysates

Analysis of CD31/PECAM1 expression in 1) Jurkat whole cell lysate, 2) human platelet lysate and 3) U937 whole cell lysate. Image from the standard format of this antibody.
Immunohistological Staining of CD31/PECAM-1 in Frozen Rabbit Heart

Immunohistological Staining of CD31/PECAM-1 in Frozen Rabbit Heart

Rabbit Heart, CD31 Stained Red. Image from verified customer review. Image from the standard format of this antibody.
Immunohistological Staining of CD31/PECAM-1 in Paraffin Embedded Human Tonsil

Immunohistological Staining of CD31/PECAM-1 in Paraffin Embedded Human Tonsil

FFPE human tonsil stained with CD31/PECAM-1 Antibody (JC/70A). Image from the standard format of this antibody.
Flow Cytometry of Jurkat Cells Stained with DyLight 550 Conjugated CD31/PECAM-1 Antibody

Flow Cytometry of Jurkat Cells Stained with DyLight 550 Conjugated CD31/PECAM-1 Antibody

A surface stain was performed on Jurkat cells with the Dylight 550 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562R, blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 5 ug/mL for 20 minutes at room temperature. Both antibodies were directly conjugated to DyLight 550.
Immunocytochemistry/Immunofluorescence Staining of CD31/PECAM-1 in HUVEC Cells

Immunocytochemistry/Immunofluorescence Staining of CD31/PECAM-1 in HUVEC Cells

HUVEC cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with CD31/PECAM-1 Antibody (JC/70A) at 2 ug/ml overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.
Immunocytochemistry/Immunofluorescence Analysis of CD31/PECAM-1 in Feline Endothelial Cells

Immunocytochemistry/Immunofluorescence Analysis of CD31/PECAM-1 in Feline Endothelial Cells

Imaging of Feline Endothelial cells with antibody dilutionof 1:25. CD31/PECAM-1 (Red), DAPI (Blue). This image was submitted via customer Review. Image from the standard format of this antibody.
Immunohistological Analysis of CD31/PECAM-1 in Paraffin Embedded Human Spleen

Immunohistological Analysis of CD31/PECAM-1 in Paraffin Embedded Human Spleen

Analysis of CD31/PECAM1 in human spleen using DAB with hematoxylin counterstain. Image from the standard format of this antibody.
Comparison of Immunohistological Staining in Human and Mouse Tissue with CD31/PECAM-1 Antibody

Comparison of Immunohistological Staining in Human and Mouse Tissue with CD31/PECAM-1 Antibody

Analysis using the Biotin conjugate of NB600-562. Comparison between mouse (lung, top) and human (prostate, bottom) staining using anti-CD31 antibody. Data demonstrates that the antibody stains human, but not mouse CD31. Image from verified customer review
Flow Cytometry of HUVEC Cells Stained with CD31/PECAM-1 Antibody

Flow Cytometry of HUVEC Cells Stained with CD31/PECAM-1 Antibody

A cell surface stain was performed on HUVEC cells with ( NB600-562, blue) along with a matched isotype control NBP2-27287 (orange). Cells were incubated in an antibody dilution of 1:100 for 20 minutes at RT. (Panel A). A negative control (HeLa cells) was also stained to ensure antibody specificity (Panel B). Image from the standard format of this antibody.
Flow Cytometry of THP-1 Cells Stained with Alexa Fluor 647 Conjugated CD31/PECAM-1 Antibody

Flow Cytometry of THP-1 Cells Stained with Alexa Fluor 647 Conjugated CD31/PECAM-1 Antibody

A surface stain was performed on THP-1 cells with the Alexa Fluor 647 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562AF647, blue) and a matched isotype control (orange). Cells were incubated in an antibody dilution of 2.5 ug/mL for 20 minutes at room temperature. Both antibodies were directly conjugated to Alexa Fluor 647.
Flow Cytometry of Jurkat Cells Stained with Alexa Fluor 488 Conjugated CD31/PECAM-1 Antibody

Flow Cytometry of Jurkat Cells Stained with Alexa Fluor 488 Conjugated CD31/PECAM-1 Antibody

A cell surface stain was performed on Jurkat cells with the Alexa Fluor 488 conjugate of CD31/PECAM-1 Antibody (JC/70A) (NB600-562AF488, blue) along with a matched isotype control. Cells were incubated in an antibody dilution of 10 ug/ml for 20 minutes at RT. Both antibodies were directly conjugated to Alexa Fluor 488.
Flow Cytometry of THP-1 Cells Stained with Alexa Fluor 488 Conjugated CD31/PECAM-1 Antibody

Flow Cytometry of THP-1 Cells Stained with Alexa Fluor 488 Conjugated CD31/PECAM-1 Antibody

A cell surface stain was performed on THP-1 cells with the Alexa Fluor 488 conjugate of CD31/PECAM-1 Antibody (JC/70A) NB600-562AF488 (blue) along with a matched isotype control. Cells were incubated in an antibody dilution of 5 ug/ml for 20 minutes at RT. Both antibodies were directly conjugated to Alexa Fluor 488.
Dual RNAscope ISH-IHC Analysis of CD31/PECAM-1 in Human Tonsil

Dual RNAscope ISH-IHC Analysis of CD31/PECAM-1 in Human Tonsil

Formalin-fixed paraffin-embedded tissue sections of human metastatic tonsil were probed for CD31 mRNA (ACD RNAScope probe, catalog # 487381; Fast Red chromogen, ACD catalog # 322500). Adjacent tissue section was processes for immunohistochemistry using mouse monoclonal (NB600-562) at 1:25 dilution for 1 hour at room temperature followed by incubation with the anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Image from the standard format of this antibody.
Knockout Validation of CD31/PECAM-1 Antibody in THP-1 Cells by Immunocytochemistry/Immunofluorescence

Knockout Validation of CD31/PECAM-1 Antibody in THP-1 Cells by Immunocytochemistry/Immunofluorescence

CD31/PECAM-1 was detected in immersion fixed THP-1 (red, positive) but not THP-1 KO (negative) cells using 8 ug/mL Mouse Anti-Human CD31/PECAM-1 (JC/70A) Monoclonal Antibody (Catalog # NB600-562) for 3 hours at room temperature. Cells were stained with the NorthernLights(TM) 557-conjugated Donkey anti-Mouse IgG Secondary Antibody (red, Catalog # NL007)) and and counterstained with DAPI (blue). Image from the standard format of this antibody.

Applications for CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free

Application
Recommended Usage

ELISA

reported in scientific literature (PMID 8307606)

Flow (Cell Surface)

1:100-1:250

Flow Cytometry

1:100-1:250

Immunohistochemistry-Paraffin

1:10-1:25

Western Blot

1:100-1:500
Application Notes
This CD31/PECAM1 Antibody (JC/70A) reacts with an ~100 kDa glycoprotein expressed by endothelial cells and ~130 kDa glycoprotein present in platelets. It stains endothelial cells in normal as well as malignant tissues. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Reviewed Applications

Read 1 review rated 1 using NBP2-80641 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 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. After opening store under sterile conditions.

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

Gene Symbol

PECAM1

Additional CD31/PECAM-1 Products

Product Documents for CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free

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.

Customer Reviews for CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free (1)

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  • CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free
    Name: Aida Meghraoui
    Application: Imaging mass cytometry
    Sample Tested: FFPE
    Species: Human
    Verified Customer | Posted 10/05/2020
    Image of human cutaneous squamous cell carcinoma tumor stained with Anti-CD31 azide/BSA free antibody [JC/70A] (in red).
    this doesn't work in IMC
    CD31/PECAM-1 Antibody (JC/70A) - Azide and BSA Free NBP2-80641
    Bio-Techne Response
    This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

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