Product Datasheet. CD31/PECAM-1 Antibody (MEC13.3) NB Unit Size: 0.1 ml

Transcription

1 Product Datasheet CD31/PECAM-1 Antibody (MEC13.3) NB Unit Size: 0.1 ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 2 Publications: 14 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 9/24/2018 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 NB CD31/PECAM-1 Antibody (MEC13.3) Product Information Unit Size Concentration Storage Clonality Clone Preservative Isotype Purity Buffer Product Description Host 0.1 ml 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Monoclonal MEC % Sodium Azide IgG2a Kappa Protein G purified PBS Rat Gene ID 5175 Gene Symbol Species Reactivity Notes Immunogen Product Application Details Applications PECAM1 Human, Mouse Mouse. mouse endothelial cell line T-end Flow Cytometry, Flow (Cell Surface), Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, In vitro assay, In vivo assay, Immunoprecipitation, CyTOF-ready Recommended Dilutions Flow Cytometry 1:10-1:1000, Immunohistochemistry 1:100-1:500, Immunocytochemistry/Immunofluorescence 1:1000-1:2000, Immunoprecipitation 1:10-1:500, Immunohistochemistry-Paraffin 1:100-1:500, Immunohistochemistry- Frozen, In vitro assay, In vivo assay, Flow (Cell Surface), CyTOF-ready Application Notes Page 1 of 6 v.20.1 Updated 9/24/2018 This CD31/PECAM1 Antibody (MEC13.3) is useful for Flow Cytometry, Immunoprecipitation, Immunohistochemical staining of acetone-fixed frozen sections and zinc-fixed paraffin-embedded sections, and in vitro and in vivo blocking of CD31-mediated cell-cell interactions. Please Note: NB works well on acetone-fixed frozen sections as well as zinc-fixed paraffinembedded sections. However, it is not recommended for formalin-fixed paraffinembedded sections due to inconsistent results. In vivo assay was reported in scientific literature. Use in vitro assays reported in scientific literature (PMID ). This antibody is CyTOF ready.

3 Images Immunocytochemistry/Immunofluorescence: CD31/PECAM-1 Antibody (MEC13.3) [NB ] - MS1 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-cd31/pecam-1 (MEC13.3) at 5.0 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. Page 2 of 6 v.20.1 Updated 9/24/2018 Immunohistochemistry-Frozen: CD31/PECAM-1 Antibody (MEC13.3) [NB ] - 4T1 tumor sections were stained for CD31 (green) and CD105 (red). Flow (Cell Surface): CD31/PECAM-1 Antibody (MEC13.3) [NB ] - A surface stain was performed on MS-1 Cells with CD31 (MEC13.3) antibody NB (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, followed by rat F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0113, R&D Systems). Flow Cytometry: CD31/PECAM-1 Antibody (MEC13.3) [NB ] - C57BL/6 mouse splenocytes stained with purified MEC13.3, followed by anti-rat IgG FITC

4 Immunocytochemistry/Immunofluorescence: CD31/PECAM-1 Antibody (MEC13.3) [NB ] - CD31/PECAM1 (MEC13.3) antibody was tested in Wehi-3 cells with Dylight 488 (green). Nuclei and beta-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Page 3 of 6 v.20.1 Updated 9/24/2018 Immunohistochemistry-Frozen: CD31/PECAM-1 Antibody (MEC13.3) [NB ] - Staining of mouse CD31 in non-small cell lung cancer tumor sections from a human NSCLC tumor-bearing mouse. This image was submitted via customer Review. Flow (Cell Surface): CD31/PECAM-1 Antibody (MEC13.3) [NB ] - A surface stain was performed on WEHI-3 Cells with CD31 (MEC13.3) antibody NB (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, followed by rat F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0113, R&D Systems).

5 Publications Abraham V, Cao G, Parambath A et al. Involvement of TIMP-1 in PECAM-1-mediated tumor dissemination Int. J. Oncol. May :00AM [PMID: ] (Human) Perales-Puchalt A, Perez-Sanz J, Payne KK et al. Frontline Science: Microbiota reconstitution restores intestinal integrity after cisplatin therapy. J. Leukoc. Biol. Mar :00AM [PMID: ] (IHC) England C, Jiang D, Hernandez R et al. ImmunoPET Imaging of CD146 in Murine Models of Intrapulmonary Metastasis of Non-Small Cell Lung Cancer. Mol Pharm 2017 Sep 06 [PMID: ] (ICC/IF, FLOW, Human) Hernandez R, England CG, Yang Y et al. ImmunoPET imaging of tissue factor expression in pancreatic cancer with (89)Zr-Df-ALT-836. J Control Release 2017 Aug 24 [PMID: ] (Mouse) DeMars KM, Pacheco SC, Yang C et al. Selective Inhibition of Janus Kinase 3 Has No Impact on Infarct Size or Neurobehavioral Outcomes in Permanent Ischemic Stroke in Mice. Front Neurol 2017 Aug 09 [PMID: ] (ICC/IF, Mouse) Takeda A, Hossain MS, Rantakari P et al. Thymocytes in Lyve1-CRE/S1pr1(f/f) Mice Accumulate in the Thymus due to Cell-Intrinsic Loss of Sphingosine-1-Phosphate Receptor Expression. Front Immunol. Nov :00AM [PMID: ] (IHC, Mouse) Salatino M, Croci DO, Laderach DJ et al. Regulation of galectins by hypoxia and their relevance in angiogenesis: strategies and methods. Methods Mol. Biol Sep 25 [PMID: ] Takeuchi K, Yanai R, Kumase F et al. EGF-Like-Domain-7 Is Required for VEGF-Induced Akt/ERK Activation and Vascular Tube Formation in an Ex Vivo Angiogenesis Assay. PLoS ONE 3/20/2014 [PMID: ] (ICC/IF, invitro, Mouse) DeLisser HM, Christofidou-Solomidou M, Strieter RM, Burdick MD, Robinson CS, Wexler RS, Kerr JS, Garlanda C, Merwin JR, Madri JA, Albelda SM. Involvement of endothelial PECAM-1/CD31 in angiogenesis. Am J Pathol;151 (3): Sep. [PMID: ] (In vivo, Mouse) Vecchi A, Garlanda C, Lampugnani MG, Resnati M, Matteucci C, Stoppacciaro A, Schnurch H, Risau W, Ruco L, Mantovani A et al. Monoclonal antibodies specific for endothelial cells of mouse blood vessels. Their application in the identification of adult and embryonic endothelium. Eur J Cell Biol;63(2): Apr. [PMID: ] Rosenblum WI, Murata S, Nelson GH, Werner PK, Ranken R, Harmon RC. Anti-CD31 delays platelet adhesion/aggregation at sites of endothelial injury in mouse cerebral arterioles. Am J Pathol;145(1): Jul. [PMID: ] (In vivo, Mouse) Baldwin HS, Shen HM, Yan HC, DeLisser HM, Chung A, Mickanin C, Trask T, Kirschbaum NE, Newman PJ, Albelda SM et al. Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31): alternatively spliced, functionally distinct isoforms expressed during mammalian cardiovascular development. Development;120(9): Sep. [PMID: ] (FLOW, Mouse) More publications at Page 4 of 6 v.20.1 Updated 9/24/2018

6 Procedures Immunocytochemistry/Immunofluorescence Protocol for CD31/PECAM1 Antibody (NB ) Immunocytochemistry Protocol Culture cells to appropriate density in 35 mm culture dishes or 6-well plates. Page 5 of 6 v.20.1 Updated 9/24/ Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes. 2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes. 3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes. 4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature. 5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature. 6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes. 7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature. 8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes. 9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing. *The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

7 Novus Biologicals USA E. Briarwood Avenue Centennial, CO USA Phone: Toll Free: Fax: Novus Biologicals Canada 461 North Service Road West, Unit B37 Oakville, ON L6M 2V5 Canada Phone: Toll Free: Fax: Novus Biologicals Europe 19 Barton Lane Abingdon Science Park Abingdon, OX14 3NB, United Kingdom Phone: (44) (0) Free Phone: Fax: (44) (0) General Contact Information Technical Support: Orders: General: Products Related to NB HAF005 Goat anti-rat IgG Secondary Antibody [HRP (Horseradish Peroxidase)] NB7115 Goat anti-rat IgG (H+L) Secondary Antibody [HRP] NBP mg Rat IgG2a Kappa Light Chain Isotype Control (R2a) NB R CD31/PECAM-1 Antibody (MEC13.3) [DyLight 550] 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. For more information on our 100% guarantee, please visit Earn gift cards/discounts by submitting a review: Earn gift cards/discounts by submitting a publication using this product: