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88560
CD20 (2H7) Mouse mAb (violetFluor™ 450 Conjugate)
Antibody Conjugates

CD20 (2H7) Mouse mAb (violetFluor™ 450 Conjugate) #88560

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Flow cytometric analysis of live human peripheral blood mononuclear cells using CD20 (2H7) Mouse mAb (violetFluor™ 450 Conjugate) (solid line) compared to concentration-matched Mouse Isotype Control (violetFluor™ 450 Conjugate) (dashed line).

To Purchase # 88560S
製品番号 サイズ 価格 在庫
88560S
500 µl  (100 tests)

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Isotype Mouse IgG2b, kappa

Product Description

This Cell Signaling Technology antibody is conjugated to violetFluor™ 450 and tested in-house for direct flow cytometric analysis in human cells.

Product Usage Information

Application Dilutions
Flow Cytometry 1:20

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.

Protocol

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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

Specificity / Sensitivity

CD20 (2H7) Mouse mAb (violetFluor™ 450 Conjugate) recognizes endogenous levels of total CD20 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Human

Source / Purification

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.

Background

B-lymphocyte antigen CD20 (also known as MS4A1; Membrane-spanning 4-domains subfamily A member 1) is a cell surface phosphoprotein involved in the regulation of B cell activation and proliferation (1,2). It is commonly used as a marker to identify B cells and is expressed throughout B cell development, up until their differentiation into plasma cells. CD20 has no known ligand, and its expression and function are largely conserved between human and mouse (1-3). Evidence suggests that CD20 is necessary for store operated calcium (SOC) entry, which leads to elevated cytoplasmic calcium levels required for B cell activation (4-5). Anti-CD20 antibody immunotherapy depletes B cells by activation of the innate monocytic network and is a common treatment for B cell lymphomas, leukemias, and autoimmune diseases (6).

The 2H7 antibody is widely used to identify both normal and malignant B cells (7).

  1. Stashenko, P. et al. (1980) J Immunol 125, 1678-85.
  2. Tedder, T.F. et al. (1985) J Immunol 135, 973-9.
  3. Tedder, T.F. et al. (1988) J Immunol 141, 4388-94.
  4. Bubien, J.K. et al. (1993) J Cell Biol 121, 1121-32.
  5. Li, H. et al. (2003) J Biol Chem 278, 42427-34.
  6. Uchida, J. et al. (2004) J Exp Med 199, 1659-69.
  7. Liu, A.Y. et al. (1987) J Immunol 139, 3521-6.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
violetFluor is a registered trademark of Tonbo Biosciences.

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