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

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).

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).

Rabbit Heart, CD31 Stained Red. Image from verified customer review.

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).

GMF-beta is involved in neovasculogenesis in human glioblastoma. A. Co-expression of GMF-beta and CD31/PECAM-1 in several tumor cells. B. The GMF-beta+/CD31+ incomplete microvessel-like structure in hypovascular zones. C. GMF-beta+/CD31+ immature microvessel in hypovascular zones. D. Single GMF-beta+/CD31+ mature microvessel in vascularized areas. E. GMF-beta-/CD31+ mature microvessels in vascularized areas. Red arrows denote GMF-beta staining, black arrows indicate CD31/PECAM-1 staining. Scale bar: 50 um. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/abstract/5509), licensed under a CC-BY licence.

Analysis of CD31/PECAM1 in human spleen using DAB with hematoxylin counterstain.

Analysis of CD31/PECAM1 expression in 1) Jurkat whole cell lysate, 2) human platelet lysate and 3) U937 whole cell lysate.

Comparison of the expression of NMDAR1 on vessels and EndoMT markers in the urinary bladder.Biopsy specimens of frozen sections from the urinary bladders of (A, C, E) normal control subjects and (B, D, F) KC patients. (A, B) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and NMDAR1 (red); DAPI was used as a nuclear stain (blue); co-expression of CD31 and NMDAR1 was evident in two groups (yellow, denoted by white arrows). (CF) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and two mesenchymal markers: FSP1 (red, in panels C and D) and -SMA (red, in panels E and F); DAPI was used as a nuclear stain (blue); co-expression of CD31 and FSP1 or -SMA was evident in three groups (yellow, denoted by white arrows) (white bars: 50 m). Microvascular Injury in Ketamine-Induced Bladder Dysfunction. PLoS One (2016)

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 conjugated to DyLight 550.

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.

Imaging of Feline Endothelial cells with antibody dilution of 1:25. CD31/PECAM-1 (Red), DAPI (Blue). This image was submitted via customer Review.

FFPE human tonsil stained with CD31/PECAM-1 Antibody (JC/70A).

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 conjugated to Alexa Fluor 488.

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 conjugated to Alexa Fluor 488.

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 conjugated to Alexa Fluor 647.

• Reactivity: Hu, Mu, Fe, Rb
• Applications: WB, ELISA, Flow, ICC/IF, IHC, Dual ISH-IHC, KO
• Clone: JC/70A
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

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

• Additional Information: Clone JC/70A was used by HLDA to establish CD designation.
• Immunogen: This CD31/PECAM-1 Antibody (JC/70A) was developed against a membrane preparation of a spleen from a patient with hairy cell leukemia.
• Localization: Cell membrane.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: PECAM1
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Dual RNAscope ISH-IHC
  • ELISA
  • Flow (Cell Surface) 1:100-1:250
  • Flow Cytometry 1:100-1:250
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:10-1:25
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:10-1:25
  • Knockout Validated
  • Western Blot 1:100-1:500
• Application Notes: This CD31/PECAM1 Antibody (JC/70A) is useful for IHC on paraffin-embedded sections, Flow cytometry and Western blot. It 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. Use in ELISA reported in scientific literature (PMID: 8307606).
• 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.

Reactivity Notes

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

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. After opening store under sterile conditions.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for CD31/PECAM-1 Antibody (JC/70A) - BSA Free

• adhesion molecule
• CD31 antigen
• CD31
• CD31/EndoCAM
• EndoCAM
• FLJ34100
• FLJ58394
• GPIIA'
• JC70A CD31
• JC70A Clone
• PECA1
• PECAM1
• PECAM-1
• PECAM-1, CD31/EndoCAM
• platelet endothelial cell adhesion molecule
• platelet endothelial cell adhesion molecule-1
• platelet/endothelial cell adhesion molecule

Background

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

Limitations

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.

Publications for CD31/PECAM-1 Antibody (NB600-562)(43)

Publications using NB600-562

• Brady EL, Prado O, Johansson F et al. Engineered tissue vascularization and engraftment depends on host model Scientific reports 2023-02-03 [PMID: 36737618] (IHC-WhMt)
• Chung S, Kim S, Lee K et al. In Situ Reprogrammings of Splenic CD11b + Cells by Nano?Hypoxia to Promote Inflamed Damage Site?Specific Angiogenesis Advanced Functional Materials 2023-05-01
• Brady E Host-Dependent Vascularization in Cardiac Tissue Engineering Thesis 1905-07-13 (WB)
• Xie C, Liang R, Ye J et al. High-efficient engineering of osteo-callus organoids for rapid bone regeneration within one month Biomaterials 2022-08-19 [PMID: 36031458]
• Xiao W, Chen W, Wang Y et al. Recombinant DT beta 4-inspired porous 3D vascular graft enhanced antithrombogenicity and recruited circulating CD93+/CD34+ cells for endothelialization Science advances 2022-07-15 [PMID: 35857526] (ICC/IF, Rat)
• Gan F, Liu L, Zhou Q et al. Effects of adipose-derived stromal cells and endothelial progenitor cells on adipose transplant survival and angiogenesis PloS one 2022-01-13 [PMID: 35025920] (ICC/IF, Rabbit)
• Yu Y, Wang X, Jia X Et al. Aptamer Probes Labeled with Lanthanide-Doped Carbon Nanodots Permit Dual-Modal Fluorescence and Mass Cytometric Imaging Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2021-11-01 [PMID: 34719883] (IHC-P, Human)
• Guzman RA, Maruyama M, Moeinzadeh S et al. The effect of genetically modified platelet-derived growth factor-BB over-expressing mesenchymal stromal cells during core decompression for steroid-associated osteonecrosis of the femoral head in rabbits Stem cell research & therapy 2021-09-15 [PMID: 34526115] (IHC-Fr, Rabbit)
• Maruyama M, Moeinzadeh S, Guzman RA et al. The efficacy of lapine preconditioned or genetically modified IL4 over-expressing bone marrow-derived mesenchymal stromal cells in corticosteroid-associated osteonecrosis of the femoral head in rabbits Biomaterials 2021-06-21 [PMID: 34186237]
• Pitale PM, Saltykova IV, Adu-Agyeiwaah Y et al. Tribbles Homolog 3 Mediates the Development and Progression of Diabetic Retinopathy Diabetes 2021-05-11 [PMID: 33975909]
• Khan M, Ashraf M Human iPS Cells Derived Skeletal Muscle Progenitor Cells Promote Myoangiogenesis and Restore Dystrophin in Duchenne Muscular Dystrophic Mice Stem Cell Res Ther 2021-02-13 [PMID: 33579366] (IHC-Fr, Human)
• Woloshuk A, Khochare S, Almulhim A et al. In situ classification of cell types in human kidney tissue using 3D nuclear staining Cytometry A 2020-11-30 [PMID: 33252180]
• Maruyama M, Lin T, Kaminow NI et al. The efficacy of core decompression for steroidassociated osteonecrosisof the femoral head in rabbits J Orthop Res 2020-10-23 [PMID: 33095462]
• Kuang XY, Ren Y, Chen C et al. Quantitative analysis for the differences in vasculogenic activity and sensitivity to angiogenic stimulants between human glioma cells and normal endothelial cells Brain Res. 2020-08-28 [PMID: 32866544] (IF/IHC, Mouse)
• Tone T, Shimizu Y, Saito H et al. In vivo behavior of untreated and compressed concentrated growth factors as biomaterials in rabbits Dent Mater J 2020-08-07 [PMID: 32779604] (IHC-P, Rabbit)
• Brady El, Kirby Ma, Olszewski E et Al. Guided vascularization in the rat heart leads to transient vessel patterning APL Bioeng. [PMID: 32161835] (IHC-P, Human)
• Xu G, Yamamoto N, Nojima T, et al. The process of bone regeneration from devitalization to revitalization after pedicle freezing with immunohistochemical and histological examination in rabbits Cryobiology 2019-12-22 [PMID: 31875528] (IF/IHC, Rabbit)
• Quintela-Fandino M, Morales S, Cortes-Salgado A, Manso L et al. Randomized Phase 0/I Trial of the Mitochondrial Inhibitor ME-344 or Placebo Added to Bevacizumab in Early HER2-Negative Breast Cancer Clin Cancer Res 2019-10-11 [PMID: 31597662] (IHC-P, Human)
• Kimball EE, Sayce L, Powell M et al. Different Vibratory Conditions Elicit Different Structural and Biological Vocal Fold Changes in an In-Vivo Rabbit Model of Phonation J Voice 2019-09-18 [PMID: 31542239] (IHC-Fr, Rabbit)
• Damond N, Engler S, Zanotelli VRT et al. A Map of Human Type 1 Diabetes Progression by Imaging Mass Cytometry Cell Metab. 2019-01-11 [PMID: 30713109] (IF/IHC, Human)
• Lee SJ, Won JE, Han C et al. Development of a three-dimensionally printed scaffold grafted with bone forming peptide-1 for enhanced bone regeneration with in vitro and in vivo evaluations J Colloid Interface Sci 2018-12-28 [PMID: 30611042] (IHC-P, Rabbit)
• Wilson C L, Stephenson S E et al. Characterization of human PDGFR-beta-positive pericytes from IPF and non-IPF lungs. Am J Physiol Lung Cell Mol Physiol 2018-01-12 [PMID: 30335500] (ICC/IF, Human)
• Miyanaga T, Ueda Y, Miyanaga A et al. Angiogenesis after administration of basic fibroblast growth factor induces proliferation and differentiation of mesenchymal stem cells in elastic perichondrium in an in vivo model: mini review of three sequential republication-abridged reports. Cell. Mol. Biol. Lett. 2018-10-05 [PMID: 30323846] (IHC-P, Rabbit)
• Hathi D, DeLassus E, Achilefu S et al. Imaging melphalan therapy response in preclinical extramedullary myeloma with 18F-FDOPA and 18F-FDG PET J. Nucl. Med. 2018-04-26 [PMID: 29700126] (Mouse)
• Vlachogiannis G, Hedayat S, Vatsiou A et al. Patient-derived organoids model treatment response of metastatic gastrointestinal cancers. Science 2018-02-23 [PMID: 29472484]
• He Y, Mei L, Jin Y, Li X. Overexpression of hepatocyte growth factor mRNA induced by gene transfer attenuates neointimal hyperplasia after balloon injury. Hum. Gene Ther. 2018-01-30 [PMID: 29382231] (IF/IHC, Rabbit)
• Ishihara T, Yamashita H, Sakurai T et al. Morphological analysis of patchy thickening and reddish discoloration of active hair growth areas in the skin of New Zealand White rabbits J Toxicol Pathol. 2017-08-18 [PMID: 29097841] (IF/IHC, Rabbit)
• Lin CC, Lin AT, Yang AH, Chen KK. Microvascular Injury in Ketamine-Induced Bladder Dysfunction. PLoS ONE. 2016-08-17 [PMID: 27529746] (ICC/IF)
• Strilic B, Yang L, Albarran-Juarez J et al. Tumour-cell-induced endothelial cell necroptosis via death receptor 6 promotes metastasis. Nature. 2016-08-11 [PMID: 27487218] (IF/IHC, Human)
• Kuang XY, Jiang XF, Chen C et al. Expressions of glia maturation factor-B by tumor cells and endothelia correlate with neovascularization and poor prognosis in human glioma. Oncotarget. 2016-12-27 [PMID: 26515590] (IF/IHC, Human)
• Song Y, Zheng J, Yan M et al. The Effects of Irreversible Electroporation on the Achilles Tendon: An Experimental Study in a Rabbit Model PLoS ONE 2015-06-27 [PMID: 26114962] (IHC-P, Rabbit)
  
  Details:
  CD31/PECAM-1 antibody (clone JC/70A) was used at 1:50 dilution for IHC-P analysis of irreversible electroporation/IRE-ablated segment, a distal segment and a proximal segment of muscle-tendon complexes from Rabbits (Fig 8).
• Sun Dongchong, Yang Yong, Wei Zhitao et al. Engineering of pre-vascularized urethral patch with muscle flaps and hypoxia-activated hUCMSCs improves its therapeutic outcome. J Cell Mol Med. 2014-01-25 [PMID: 24460735] (IF/IHC)
• Song X, Wang J, Zheng T et al. LBH589 Inhibits proliferation and metastasis of hepatocellular carcinoma via inhibition of gankyrin/stat3/akt pathway. Mol Cancer. 2013-10-05 [PMID: 24093956] (WB, Human)
• Kalen M, Heikura T, Karvinen H et al. Gamma-secretase inhibitor treatment promotes VEGF-A-driven blood vessel growth and vascular leakage but disrupts neovascular perfusion. PLoS One. 2011-04-14 [PMID: 21533193] (IF/IHC, Rabbit)
• Longatto Filho A, Oliveira TG, Pinheiro C et al. How useful is the assessment of lymphatic vascular density in oral carcinoma prognosis? World J Surg Oncol. 2007-12-11 [PMID: 18072963] (IF/IHC, Human)
• Ding MX, Lin XQ, Fu XY Zhang N, Li JC. Expression of vascular endothelial growth factor-C and angiogenesis in esophageal squamous cell carcinoma World J Gastroenterol. 2006-07-28 [PMID: 16874878] (IF/IHC, Human)
• Elaqoz S, Eqilmez R, Koyuncu A et al. The intratumoral microvessel density and expression of bFGF and nm23-H1 in colorectal cancer. Pathol Oncol Res. 2006-01-01 [PMID: 16554912] (IF/IHC, Human)
• Reymond N, Imbert AM, Devilard E et al. DNAM-1 and PVR regulate monocyte migration through endothelial junctions. J Exp Med. 2004-05-17 [PMID: 15136589] (FLOW, IF/IHC, Human)
• Zurita AJ, Troncoso P, Cardo-Vila M et al. Combinatorial screenings in patients: the interleukin-11 receptor alpha as a candidate target in the progression of human prostate cancer. Cancer Res. 2004-01-15 [PMID: 14744752] (IF/IHC, Human)
• Hambrecht R, Adams V, Erbs S. Regular physical activity improves endothelial function in patients with coronary artery disease by increasing phosphorylation of endothelial nitric oxide synthase. Circulation. 2003-07-01 [PMID: 12810615] (IF/IHC, Human)
• Ikpeazu C, Davidson MK, halteman D, et al. Donor origin of circulating endothelial progenitors after allogeneic bone marrow transplantation. Biol Blood Marrow Transplant. 2000-01-01 [PMID: 10905767] (IF/IHC, Human)
• Tjalma W, Van Marck E, Weyler J et al. Quantification and prognostic relevance of angiogenic parameters in invasive cervical cancer. Br J Cancer. 1998-07-01 [PMID: 9683289] (IF/IHC, Human)
• Bird IN, Spragg JH, Ager A, Matthews N. Studies of lymphocyte transendothelial migration: analysis of migrated cell phenotypes with regard to CD31 (PECAM-1), CD45RA and CD45RO. Immunology. 1993-12-01 [PMID: 8307606] (ELISA, Human)

Reviews for CD31/PECAM-1 Antibody (NB600-562) (4)

Average Rating: 4.3

(Based on 4 reviews)

We have 4 reviews tested in 3 species: Mouse, Feline, Other.

reviewed by: Kirsten Hattermann-Koch IHC-Fr Mouse 05/09/2019

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Mouse brain
• Species: Mouse
• Lot: D

reviewed by: Kazuya Oikawa ICC Feline 05/26/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Endothelial
• Species: Feline
• Lot: C-1

reviewed by: Shrindih Joshi IHC-P Mouse 08/12/2016

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: non-ischemic and ischemic gastrocnemius muscle
• Species: Mouse
• Lot: A

reviewed by: Alejandra Casar IHC-Fr Other 04/28/2014

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Rabbit Heart
• Species: Other
• Lot: A-1

FAQs for CD31/PECAM-1 Antibody (NB600-562). (Showing 1 - 3 of 3 FAQs).

1. I have an inquiry about #NB600-562, Anti-CD31/PECAM1 (JC/70A), and wanted to know the recommended IHC-P antigen retrieval protocol?
   • Please see our protocols for antigen retrieval. There are different methods of antigen retrieval as you will see; our lab typically recommends the heat induced citrate buffer pH 6.0 antigen retrieval method as the most commonly used in our lab.
2. What's the possibility of using this antibody for IHC with frozen sections?
   • Although this product has only been confirmed for use in IHC-P, it most likely will work with frozen sections as well.
3. Would this antibody react with samples from Rabbit or not? Because only Hu is listed as its Species, but it is also mentioned that Cross-reacts with Human and Rabbit.
   • Yes, this antibody should react with rabbit and will be covered as such in our 100% guarantee.

Bioinformatics

• Gene Symbol: PECAM1
• Entrez:
  • 5175(Human)
• Uniprot:
  • P16284()