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Immunofluorescent analysis of P-selectin/CD62P was performed on 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ P-selectin/CD62P recombinant rabbit monoclonal antibody (Product # 701257) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 113–126 of human P-selectin|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with mouse, rat and non-human primate based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
P-selectin is a 140 kDa type 1 transmembrane glycoprotein, and is one of the most commonly studied proteins that regulate cell rolling. P-selectin is stored in the Weibel-Palade bodies of endothelial cells, as well as in a-granules of platelets. From there, it can be rapidly brought to the cell surface after exposure to thrombin, histamine, complement 5a, Ca21 ionophores, or adenosine diphosphate.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
CD62; CD62 antigen-like family member P; CD62P; FLJ45155; GMP-140; GMP140; granule membrane protein 140; granule membrane protein 140kDa; granulocyte membrane protein; GRMP; LECAM3; leukocyte-endothelial cell adhesion molecule 3; PADGEM; platelet activation dependent granule-external membrane protein; platelet alpha-granule membrane protein; PSEL; RP1-86F14.2; selectin P (granule membrane protein 140kDa, antigen CD62)
CD62; CD62P; GMP140; GMRP; GRMP; LECAM3; PADGEM; PSEL; SELP