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27221
CD19 (1D3) Rat mAb (PE-Cy7® Conjugate)
Antibody Conjugates

CD19 (1D3) Rat mAb (PE-Cy7® Conjugate) #27221

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
M Endogenous Rat IgG2a, kappa
Flow Cytometry

Flow cytometric analysis of live mouse splenocytes using CD19 (1D3) Rat mAb (PE-Cy7® Conjugate) (solid line) compared to concentration-matched Rat Isotype Control (PE-Cy7® Conjugate) (dashed line).

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Flow Cytometry

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

For optimal flow cytometry results, we recommend 0.25 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:80
Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

CD19 (1D3) Rat mAb (PE-Cy7® Conjugate) recognizes endogenous levels of total mouse CD19 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

This Cell Signaling Technology antibody is conjugated to PE-Cy7® and tested in-house for direct flow cytometric analysis in mouse cells.

CD19 is a 95 kDa coreceptor, which amplifies the signaling cascade in B cells (1). On the B cell surface, CD19 associates with CD21, CD81 and Leu-13 to exert its function. The cytoplasmic tail of CD19 has nine conserved tyrosine residues playing critical roles in CD19 mediated function by coupling signaling molecules to the receptor (1). After B cell receptor or CD19 ligation, Tyr531 and Tyr500 of CD19 are progressively phosphorylated. This phosphorylation enables the coupling of PI3 kinase and Src family tyrosine kinase to CD19 and activates the PI3K and Src signaling pathways (2,3). Coligation of B cell receptor and CD19 also promotes Tyr409 phosphorylation in CD19. The phosphorylation at these sites enables its binding to Vav and mediates elevated intracellular calcium response, as well as the JNK pathway (4,5).

The 1D3 clone is a widely used as a phenotypic marker for CD19 expression on the surface of murine B cells and subsets of dendritic cells.

  1. Tedder, T.F. et al. (1997) Immunity 6, 107-18.
  2. Buhl, A.M. and Cambier, J.C. (1999) J Immunol 162, 4438-46.
  3. Fujimoto, M. et al. (2000) Immunity 13, 47-57.
  4. O'Rourke, L.M. et al. (1998) Immunity 8, 635-45.
  5. Sato, S. et al. (1997) Proc Natl Acad Sci U S A 94, 13158-62.
Entrez-Gene Id
12478
Swiss-Prot Acc.
P25918
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Cy and CyDye are registered trademarks of GE Healthcare

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To Purchase # 27221S

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Product # Size Price
27221S
100 µg $ 199.0