FineTest®Violet 450 Anti-Mouse CD162 Antibody(4RA10)

Synonyms:CD162|P-selectin glycoprotein ligand 1|PSGL-1|Selectin P ligand|Selplg|PSGL1 antibody
Catalogue No.:FV450-30280Reactivity:Mouse
Host:RatTested Application:FC
Clonality:MonoclonalIsotype:IgG1, κ
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Product Name
FineTest®Violet 450 Anti-Mouse CD162 Antibody(4RA10)
Catalogue No.
FV450-30280
Form
liquid
Conjugation
FineTest®Violet 450
Clonality
Monoclonal
Isotype
IgG1, κ
Clone ID
4RA10
Storage
PBS with 0. 1% sodium azide, 1%BSA, pH 7.3, 2-8℃ for 12 months (Avoid repeated freeze / thaw cycles.)
Immunogen
Alternative Names
CD162|P-selectin glycoprotein ligand 1|PSGL-1|Selectin P ligand|Selplg|PSGL1 antibody
UniProt ID
Q62170
Application
Tested Applications
FC
Recommended dilution
Volume per test: 5μL. Each lot of this antibody is quality control tested by flow cytometric analysis. The amount of the reagent is suggested to be used 5 µL of antibody per test (million cells in 100 µL staining volume or per 100 µL of whole blood). Please check your vial before the experiment. Since applications vary, the appropriate dilutions must be determined for individual use.
Background
The 4RA10 antibody reacts with the N-terminal functional peptide of CD162 (P-selectin glycoprotein ligand-1, PSGL-1), encoded by the Selpl gene. PSGL-1 is expressed on the cell surface as a homodimer of approximately 230 kDa. In the mouse, Selpl mRNA is detected in most tissues, with high levels found in hematopoietic cells, brain, and adipose tissue. Flow cytometric analyses have revealed CD162 expression on bone marrow-derived mast and dendritic cells, splenic leukocytes, platelets, peripheral blood neutrophils, and neutrophil and T-cell lines. PSGL-1 is a ligand for P-selectin (CD62P) and is involved in leukocyte rolling, the migration of leukocytes into inflamed tissues, and responses to vascular injury. It is a sialomucin that must be specifically sialylated, fucosylated, and sulfated to bind P-selectin. There is also evidence that other ligands for PSGL-1 and CD62P may exist. 4RA10 mAb is reported to block the binding of mouse leukocytes to CD62P and CD62L.
How many times can antibodies be recycled?

First, usually it's not suggested to recycle antibodies. After use, buffer system of antibodies has changed. The storage condition of recycled antibodies for different customers also varies. Thus, the performance efficiency of recycled antibodies can’t be guaranteed. Besides, FineTest ever conducted the antibody recycling assay. Assay results show recycling times of different antibodies also varies. Usually, higher antibody titer allows more repeated use. Customers can determine based on experimental requirements.

Notes: After incubation, we recycle rest antibodies to centrifuge tube and store at 4℃. High titer antibodies can be stored for a minimum of one week. Reuse about three times.

What are components of FineTest antibody buffer?

Components of FineTest antibody buffer are usually PBS with proclin300 or sodium azide, BSA, 50% glycerol. Common preservative is proclin300 or sodium azide, which is widely applied in the lab and industry.

How about the storage temperature and duration of FineTest antibodies?

Most antibodies are stored at -20℃. Directly-labeled flow cytometry antibodies should be stored at 2 - 8℃. The shelf life is one year. If after sales issues for purchased antibodies appear, return or replacement is available. Usually, antibodies can be still used after the one-year warranty. We can offer technical support services.

Is dilution required for FineTest antibodies? What’s the dilute solution?

Directly-labeled flow cytometry antibodies are ready-to-use without dilution. Other antibodies are usually concentrated. Follow the dilution ratio suggested in the manual. Dilute solution for different experiments also varies. Common antibody dilution buffers are acceptable(e.g. PBST, TBST, antibody blocking buffer).

How to retrieve antibodies for immunohistochemistry?

Common retrieval buffers: Tris-EDTA Buffer(pH 9.0); Citrate Buffer(pH 6.0)

Heat induced antibody retrieval:

Method 1: Water-bath heating: Put the beaker with retrieval buffer and slide in the boiling water bath. Keep the boiling state for 15min. Naturally cool to room temperature;

Method 2: Microwave retrieval: Put the beaker with retrieval buffer and slide in the microwave oven. Heat at high power for 5min, Switch OFF for 3min, Heat at medium power for 5min. Naturally cool to room temperature.

How to choose secondary antibodies?

(1) Secondary antibodies react with primary antibodies. Thus, secondary antibodies should be against host species of primary antibodies. E.g. If the primary antibody is derived from rabbit, the relevant secondary antibody should be against rabbit. E.g. goat anti rabbit or donkey anti rabbit.

(2) Choose secondary antibody conjugates according to the experimental type, e.g. ELISA, WB, IHC etc. Common enzyme conjugated secondary antibodies are labelled by HRP, AP etc. Fluorescin or dye labelled secondary antibodies are applied in immunofluorescence and flow cytometry(e.g. FITC, Cy3).

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