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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, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

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

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com/flowdyes for a listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.

NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised August 2019

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.