P-Selectin/CD62P Antibody [Unconjugated]

Western blot shows lysates of mouse platelets and rat platelets. PVDF membrane was probed with 2 µg/mL of Goat Anti-Mouse/Rat P-Selectin/CD62P Antigen Affinity-purified Polyclonal Antibody (Catalog # AF737) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for P-Selectin/CD62P at approximately 140-150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

P‑Selectin/CD62P was detected in perfusion fixed frozen sections of mouse liver using Goat Anti-Mouse/Rat P‑Selectin/CD62P Antigen Affinity-purified Polyclonal Antibody (Catalog # AF737) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to endothelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of rat platelets, loaded at 0.2 mg/mL. A specific band was detected for P‑Selectin/CD62P at approximately 197 kDa (as indicated) using 20 µg/mL of Goat Anti-Mouse P‑Selectin/CD62P Antigen Affinity-purified Polyclonal Antibody (Catalog # AF737) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12‑230 kDa separation system.

In-vivo targeting of BYL719-loaded nanoparticles prepared with either fucoidan (Fi) or dextran sulfate (Dex).(a) Representative ex vivo fluorescence images of mice organs 24 h after i.v. administration of FiBYL719 or DexBYL719 nanoparticles, and pre-treated with anti-P-selectin antibody (Ab). (b) Nanoparticle biodistribution in organs and tumour, calculated from ex vivo fluorescence images shown in a as total fluorescence efficiency divided by organ weight (n=3). (c) Representative immunofluorescence staining for CD-31 (red), P-selectin (green) and DAPI (blue) in Cal-33 xenografts before and after a single dose of ionizing radiation (4 Gy). Scale bars, 50 μm. (d) Quantification of double-staining positive endothelial cells per tumour shown in c (n=3). (e) In vivo fluorescence imaging of Cal-33 xenograft-bearing mice 24 h after treatment with FiBYL719 or 4 Gy RT followed by FiBYL719. (f) Quantification of total fluorescence efficiency of tumours shown e (n=10). (g) Representative immunofluorescence stains of tumour sections for P-selectin (green), NIR (red) and DAPI (blue) from H22 xenografts 24 h after treatments. Scale bars, 50 μm. In b,d,f error bars indicate mean±s.e.m. *P<0.05, **P<0.01; by Mann–Whitney U-test in b or one-way ANOVA with post hoc Tukey test in d,f. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms14292), licensed under a CC-BY license. Not internally tested by R&D Systems.

In-vivo targeting of BYL719-loaded nanoparticles prepared with either fucoidan (Fi) or dextran sulfate (Dex).(a) Representative ex vivo fluorescence images of mice organs 24 h after i.v. administration of FiBYL719 or DexBYL719 nanoparticles, and pre-treated with anti-P-selectin antibody (Ab). (b) Nanoparticle biodistribution in organs and tumour, calculated from ex vivo fluorescence images shown in a as total fluorescence efficiency divided by organ weight (n=3). (c) Representative immunofluorescence staining for CD-31 (red), P-selectin (green) and DAPI (blue) in Cal-33 xenografts before and after a single dose of ionizing radiation (4 Gy). Scale bars, 50 μm. (d) Quantification of double-staining positive endothelial cells per tumour shown in c (n=3). (e) In vivo fluorescence imaging of Cal-33 xenograft-bearing mice 24 h after treatment with FiBYL719 or 4 Gy RT followed by FiBYL719. (f) Quantification of total fluorescence efficiency of tumours shown e (n=10). (g) Representative immunofluorescence stains of tumour sections for P-selectin (green), NIR (red) and DAPI (blue) from H22 xenografts 24 h after treatments. Scale bars, 50 μm. In b,d,f error bars indicate mean±s.e.m. *P<0.05, **P<0.01; by Mann–Whitney U-test in b or one-way ANOVA with post hoc Tukey test in d,f. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms14292), licensed under a CC-BY license. Not internally tested by R&D Systems.

In-vivo targeting of BYL719-loaded nanoparticles prepared with either fucoidan (Fi) or dextran sulfate (Dex).(a) Representative ex vivo fluorescence images of mice organs 24 h after i.v. administration of FiBYL719 or DexBYL719 nanoparticles, and pre-treated with anti-P-selectin antibody (Ab). (b) Nanoparticle biodistribution in organs and tumour, calculated from ex vivo fluorescence images shown in a as total fluorescence efficiency divided by organ weight (n=3). (c) Representative immunofluorescence staining for CD-31 (red), P-selectin (green) and DAPI (blue) in Cal-33 xenografts before and after a single dose of ionizing radiation (4 Gy). Scale bars, 50 μm. (d) Quantification of double-staining positive endothelial cells per tumour shown in c (n=3). (e) In vivo fluorescence imaging of Cal-33 xenograft-bearing mice 24 h after treatment with FiBYL719 or 4 Gy RT followed by FiBYL719. (f) Quantification of total fluorescence efficiency of tumours shown e (n=10). (g) Representative immunofluorescence stains of tumour sections for P-selectin (green), NIR (red) and DAPI (blue) from H22 xenografts 24 h after treatments. Scale bars, 50 μm. In b,d,f error bars indicate mean±s.e.m. *P<0.05, **P<0.01; by Mann–Whitney U-test in b or one-way ANOVA with post hoc Tukey test in d,f. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms14292), licensed under a CC-BY license. Not internally tested by R&D Systems.

Expression of P-selectin in different brain regions following GMH. P14 brain sections stained for P-selectin (CD62p) and quantified in 3 specific brain regions of each experimental group as indicated (a, b, c). One-way ANOVA with Turkey’s correction for multiple comparisons. ****p < 0.0001. Error bars = mean ± SEM. d 40 × representative images of P-selectin staining (green) and DAPI (blue) within each brain region for naïve and GMH animals Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/37322469), licensed under a CC-BY license. Not internally tested by R&D Systems.

Expression of P-selectin in different brain regions following GMH. P14 brain sections stained for P-selectin (CD62p) and quantified in 3 specific brain regions of each experimental group as indicated (a, b, c). One-way ANOVA with Turkey’s correction for multiple comparisons. ****p < 0.0001. Error bars = mean ± SEM. d 40 × representative images of P-selectin staining (green) and DAPI (blue) within each brain region for naïve and GMH animals Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/37322469), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu, Rt
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

P-Selectin/CD62P Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse P-Selectin/CD62P
  Trp42-Ala709
  Accession # Q01102
• Specificity: Detects mouse P-Selectin/CD62P in direct ELISAs. Detects mouse and rat P-Selectin/CD62P in Western blot. In direct ELISAs, less than 15% cross-reactivity with recombinant human P-Selectin/CD62P is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SELP
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for P-Selectin/CD62P Antibody [Unconjugated]

• CD62P antigen
• CD62P
• FLJ45155
• GMP140
• GRMP
• PADGEM
• PADGEMantigen CD62)
• PSEL
• P-Selectin
• selectin P (granule membrane protein 140kDa, antigen CD62)
• SELP

Background

Mouse P-Selectin (GMP-140, LECAM-3, PADGEM, CD62P), a member of the Selectin family, is a cell surface glycoprotein expressed by activated platelets and endothelial cells. P-Selectin is translocated to the cell surface within minutes, from alpha granules of platelets or Weibel-Palade bodies of endothelial cells, following stimulation with thrombin, histamine, PMA or peroxides. P-Selectin binds to a 106 kDa protein present on myeloid cells, neutrophils, monocytes and lymphocytes, termed PSGL-1 (P-Selectin glycoprotein ligand-1). P-Selectin plays a role in the adhesion of leukocytes and neutrophils to the endothelium. Acting in cooperation with L-Selectin, P-Selectin mediates the initial interaction of circulating leukocytes with endothelial cells that produces a characteristic ‘rolling’ of the leukocytes on the endothelium. This initial interaction is followed by a stronger interaction involving E-Selectin, and later ICAM-1 and VCAM-1, that leads eventually to extravasation of the white blood cell through the blood vessel wall into the extracellular matrix tissue. Mouse P-Selectin cDNA encodes a 768 amino acid (aa) residue type I transmembrane protein with a 41 aa signal peptide, a 668 aa extracellular domain, a transmembrane domain and a short (35 aa) cytoplasmic domain. The extracellular domain has an NH 2 -terminal C-type lectin domain and an EGF-like domain followed by a series of complement factor A repeat homology domains. The extracellular domains of human and mouse P-Selectin share approximately 73% sequence homology.

1. Kansas, G.S. (1996) Blood 88:3259.
2. McEver, R.P. and R.D. Cummings (1997) J. Clin. Invest. 100:485.

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 P-Selectin/CD62P Antibody (AF737)(25)

Publications using AF737

• Tranter, JD;Mikami, RT;Kumar, A;Brown, G; LRRC8 complexes are adenosine nucleotide release channels regulating platelet activation and arterial thrombosis bioRxiv 2024-01-01 [PMID: 39386563] (Western Blot, Mouse)
• Levard, D;Seillier, C;Bellemain-Sagnard, M;Fournier, AP;Lemarchand, E;Dembech, C;Riou, G;McDade, K;Smith, C;McQuaid, C;Montagne, A;Amann, L;Prinz, M;Vivien, D;Rubio, M; Central nervous system-associated macrophages modulate the immune response following stroke in aged mice Nature neuroscience 2024-07-03 [PMID: 38961228] (Immunohistochemistry, Mouse)
• CP Camargo, AK Muhuri, Y Alapan, LF Sestito, M Khosla, MP Manspeaker, AS Smith, CM Paulos, SN Thomas A dhesion analysis via a tumor vasculature-like microfluidic device identifies CD8+ T�cells with enhanced tumor homing to improve cell therapy Cell Reports, 2023-02-26;42(3):112175. 2023-02-26 [PMID: 36848287] (IHC, Mouse)
• P Marona, J Górka, O Kwapisz, J Jura, J Rys, RM Hoffman, K Miekus Resistance to tyrosine kinase inhibitors promotes renal cancer progression through MCPIP1 tumor-suppressor downregulation and c-Met activation Cell Death & Disease, 2022-09-22;13(9):814. 2022-09-22 [PMID: 36138026] (IHC, Mouse)
• Takeshi Hirata, Takahide Itokazu, Atsushi Sasaki, Fuminori Sugihara, Toshihide Yamashita Humanized Anti-RGMa Antibody Treatment Promotes Repair of Blood-Spinal Cord Barrier Under Autoimmune Encephalomyelitis in Mice Frontiers in Immunology 6/15/2022 [PMID: 35784362]
• H Li, Z Wang, Z Chen, T Ci, G Chen, D Wen, R Li, J Wang, H Meng, R Bryan Bell, Z Gu, G Dotti, Z Gu Disrupting tumour vasculature and recruitment of aPDL1-loaded platelets control tumour metastasis Nature Communications, 2021-05-13;12(1):2773. 2021-05-13 [PMID: 33986264] (Flow Cytometry, IHC, Mouse)
• Neil Dani, Rebecca H. Herbst, Cristin McCabe, Gilad S. Green, Karol Kaiser, Joshua P. Head et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages Cell 5/1/2021 [PMID: 33932339]
• Joan D. Beckman, Fuad Abdullah, Chunsheng Chen, Rachel Kirchner, Dormarie Rivera-Rodriguez, Zachary M. Kiser et al. Endothelial TLR4 Expression Mediates Vaso-Occlusive Crisis in Sickle Cell Disease Frontiers in Immunology 1/19/2021 [PMID: 33542720]
• Z Li, EA Korhonen, A Merlini, J Strauss, E Wihuri, H Nurmi, S Antila, J Paech, U Deutsch, B Engelhardt, S Chintharla, GY Koh, A Flügel, K Alitalo Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS J. Clin. Invest., 2020-04-01;0(0):. 2020-04-01 [PMID: 32149735] (IHC-P, Mouse)
• A Drieu, A Lanquetin, D Levard, M Glavan, F Campos, A Quenault, E Lemarchand, M Naveau, AL Pitel, J Castillo, D Vivien, M Rubio Alcohol exposure-induced neurovascular inflammatory priming impacts ischemic stroke and is linked with brain perivascular macrophages JCI Insight, 2020-02-27;0(0):. 2020-02-27 [PMID: 31990687] (IHC, In Vivo, Mouse)
• Yearin Jun, Jun Xu, Hyungjun Kim, Ji Eun Park, Yoo-Seong Jeong, Jee Sun Min et al. Carfilzomib Delivery by Quinic Acid-Conjugated Nanoparticles: Discrepancy Between Tumoral Drug Accumulation and Anticancer Efficacy in a Murine 4T1 Orthotopic Breast Cancer Model Journal of Pharmaceutical Sciences 4/1/2020 [PMID: 31945310]
• Gloria Vegliante, Daniele Tolomeo, Antoine Drieu, Marina Rubio, Edoardo Micotti, Federico Moro et al. Longitudinal Molecular Magnetic Resonance Imaging of Endothelial Activation after Severe Traumatic Brain Injury Journal of Clinical Medicine 7/30/2019 [PMID: 31366109]
• G Xu, C Li, AL Parsiola, J Li, KD McCarter, R Shi, WG Mayhan, H Sun Dose-Dependent Influences of Ethanol on Ischemic Stroke: Role of Inflammation Front Cell Neurosci, 2019-02-12;13(0):6. 2019-02-12 [PMID: 30853895] (Western Blot, Mouse)
• Gregory M. Vercellotti, Agustin P. Dalmasso, Terry R. Schaid, Julia Nguyen, Chunsheng Chen, Marna E. Ericson et al. Critical role of C5a in sickle cell disease American Journal of Hematology 3/1/2019 [PMID: 30569594]
• Gregory M. Vercellotti, Agustin P. Dalmasso, Terry R. Schaid, Julia Nguyen, Chunsheng Chen, Marna E. Ericson et al. Critical role of C5a in sickle cell disease American Journal of Hematology 2019-03-01 [PMID: 30569594] (Immunohistochemistry, Mouse)
• Jun Xu, Steve Seung‐Young Seung-Young Lee, Howon Seo, Liang Pang, Yearin Jun, Ruo‐Yu Zhang et al. Quinic Acid‐Conjugated Nanoparticles Enhance Drug Delivery to Solid Tumors via Interactions with Endothelial Selectins Small 12/1/2018 [PMID: 30411856]
• Roberto Tinoco, Florent Carrette, Monique L. Henriquez, Yu Fujita, Linda M. Bradley Fucosyltransferase Induction during Influenza Virus Infection Is Required for the Generation of Functional Memory CD4+ T Cells The Journal of Immunology 2018-04-15 [PMID: 29491007] (Flow Cytometry, Immunohistochemistry, Transgenic Mouse)
• Roberto Tinoco, Florent Carrette, Monique L. Henriquez, Yu Fujita, Linda M. Bradley Fucosyltransferase Induction during Influenza Virus Infection Is Required for the Generation of Functional Memory CD4+ T Cells The Journal of Immunology 4/15/2018 [PMID: 29491007]
• AP Fournier, A Quenault, S Martinez d, M Gauberti, G Defer, D Vivien, F Docagne, R Macrez Prediction of disease activity in models of multiple sclerosis by molecular magnetic resonance imaging of P-selectin Proc. Natl. Acad. Sci. U.S.A., 2017-05-22;0(0):. 2017-05-22 [PMID: 28533365] (IHC, Mouse)
• A Mizrachi, Y Shamay, J Shah, S Brook, J Soong, VK Rajasekhar, JL Humm, JH Healey, SN Powell, J Baselga, DA Heller, A Haimovitz-, M Scaltriti Tumour-specific PI3K inhibition via nanoparticle-targeted delivery in head and neck squamous cell carcinoma Nat Commun, 2017-02-13;8(0):14292. 2017-02-13 [PMID: 28194032] (IHC, Mouse)
• Aurélien Quenault Molecular magnetic resonance imaging discloses endothelial activation after transient ischaemic attack Brain, 2016-11-08;140(0):146-157. 2016-11-08 [PMID: 28031221] (Functional Assay, IHC, Mouse)
• Yosi Shamay, Moshe Elkabets, Hongyan Li, Janki Shah, Samuel Brook, Feng Wang et al. P-selectin is a nanotherapeutic delivery target in the tumor microenvironment Science Translational Medicine 6/29/2016 [PMID: 27358497]
• Clerin V, Shih HH, Deng N, Hebert G, Resmini C, Shields KM, Feldman JL, Winkler A, Albert L, Maganti V, Wong A, Paulsen JE, Keith JC, Vlasuk GP, Pittman DD Expression of the cysteine protease legumain in vascular lesions and functional implications in atherogenesis. Atherosclerosis, 2008-02-21;0(0):. 2008-02-21 [PMID: 18377911] (ICC, Mouse)
• Tikellis C, Jandeleit-Dahm KA, Sheehy K, Murphy A, Chin-Dusting J, Kling D, Sebokova E, Cooper ME, Mizrahi J, Woollard KJ Reduced plaque formation induced by rosiglitazone in an STZ-diabetes mouse model of atherosclerosis is associated with downregulation of adhesion molecules. Atherosclerosis, 2008-02-21;199(1):55-64. 2008-02-21 [PMID: 18093596] (IHC-P, Mouse)
• Washington AV, Schubert RL, Quigley L, Disipio T, Feltz R, Cho EH, McVicar DW A TREM family member, TLT-1, is found exclusively in the alpha-granules of megakaryocytes and platelets. Blood, 2004-04-20;104(4):1042-7. 2004-04-20 [PMID: 15100151] (ICC, Human)

Bioinformatics

• Gene Symbol: SELP
• Uniprot:
  • Q01102()