14-0628-37

antibody from Invitrogen Antibodies

Targeting: SELP CD62, CD62P, GMP140, GRMP, PADGEM, PSEL

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Recommended by provider)

Other assay (Recommended by provider)

Antibody Data

Product number

14-0628-37

Provider

Invitrogen Antibodies

Product name

CD62P (P-Selectin) Monoclonal Antibody (AK-4), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The AK-4 monoclonal antibody reacts with human CD62P, a 140 kDa transmembrane glycoprotein also known as P-Selectin. Upon activation, CD62P is rapidly transported from its cytoplasmic stores in the Weibel-Palade bodies of endothelial cells or the alpha-granules of platelets to the surface. CD62P initiates the adhesive interaction between endothelial cells and neutrophils and monocytes during the inflammatory reaction and is also involved in the interaction of platelets with monocytes and neutrophils. Applications Reported: The AK-4 antibody has been reported for use in flow cytometric analysis, and immunohistochemical staining of frozen tissue sections. AK-4 has also been reported in blocking adhesion of platelets in in vitro assays. Applications Tested: This AK-4 antibody has been tested by flow cytometric analysis of resting and thrombin-activated human platelets. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

AK-4

Vial size

2 mg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Soluble Siglec-5 associates to PSGL-1 and displays anti-inflammatory activity.

Pepin M, Mezouar S, Pegon J, Muczynski V, Adam F, Bianchini EP, Bazaa A, Proulle V, Rupin A, Paysant J, Panicot-Dubois L, Christophe OD, Dubois C, Lenting PJ, Denis CV
Scientific reports 2016 Nov 28;6:37953

---

Monitoring receptor-ligand interactions between surfaces by thermal fluctuations.

Chen W, Evans EA, McEver RP, Zhu C
Biophysical journal 2008 Jan 15;94(2):694-701

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of P-Selectin was performed using HEL 92.1.7 and Raji cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with P-selectin Mouse Monoclonal Antibody (Product # 14-0628-80) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing membrane localization of P-Selectin in HEL 92.1.7 but not in Raji (Panel e). Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of thrombin-activated human peripheral blood platelets with 0.5 µg of Mouse IgG1 kappa Isotype Control Purifed (Product # 14-4714-82) (open histogram) or 0.5 µg of Anti-Human CD62P (P-Selectin) Purified (filled histogram) followed by Anti-Mouse IgG FITC (Product # 11-4011-85). Total viable cells were used for analysis.