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9705
CD24 (M1/69) Rat mAb (violetFluor™ 450 Conjugate)
Antibody Conjugates

CD24 (M1/69) Rat mAb (violetFluor™ 450 Conjugate) #9705

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Flow cytometric analysis of live mouse splenocytes using CD24 (M1/69) Rat mAb (violetFluor™ 450 Conjugate)

(solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (violetFluor™ 450 Conjugate)

#52572 (dashed line).

To Purchase # 9705S
Product # Size Price
9705S
100 µg $ 279

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Isotype Rat IgG2b, kappa

Product Description

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

Product Usage Information

For optimal flow cytometry results, we recommend 0.5 μg of antibody per test.

Application Dilutions
Flow Cytometry 1:40

Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. 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

CD24 (M1/69) Rat mAb (violetFluor™ 450 Conjugate) recognizes endogenous levels of total CD24 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

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

CD24, also know as heat stable antigen HSA, is a P-selectin ligand involved in adhesion. It is a GPI-anchored glycoprotein expressed on many types of cells, including hematopoietic cells, neural cells, and epithelial cells. CD24 is widely used to delineate stages of lymphocyte development (1-3). It also binds to Siglec-10 in humans or Siglec-G in mice (4,5). CD24 is frequently used as a marker to identify and isolate cancer stem cells in various cancer types (6,7).

  1. Sammar, M. et al. (1994) Int Immunol 6, 1027-36.
  2. Aigner, S. et al. (1997) Blood 89, 3385-95.
  3. Fang, X. et al. (2010) Cell Mol Immunol 7, 100-3.
  4. Chen, G.Y. et al. (2009) Science 323, 1722-5.
  5. Toubai, T. et al. (2014) Blood 123, 3512-23.
  6. Jaggupilli, A. and Elkord, E. (2012) Clin Dev Immunol 2012, 708036.
  7. Yu, Z. et al. (2012) Int J Biochem Cell Biol 44, 2144-51.

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.