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53289
CD44 (IM7) Rat mAb (violetFluor™ 450 Conjugate)
Antibody Conjugates

CD44 (IM7) Rat mAb (violetFluor™ 450 Conjugate) #53289

APPLICATIONS

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

Flow cytometric analysis of live human peripheral blood mononuclear cells using CD44 (IM7) Rat mAb (violetFluor™ 450 Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (violetFluor™ 450 Conjugate) #52572 (dashed line).

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Flow Cytometry

Flow cytometric analysis of live mouse splenocytes using CD44 (IM7) Rat mAb (violetFluor™ 450 Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (violetFluor™ 450 Conjugate) #52572 (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 0.25μg 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, pH7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

CD44 (IM70) Rat mAb (violetFluor™ 450 Conjugate) recognizes endogenous levels of total CD44 protein. This antibody detects an epitope within the extracellular domain and is expected to detect all isoforms of CD44.

Species Reactivity:

Human, 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 violetFluor™ 450 and tested in-house for direct flow cytometric analysis in human and mouse cells.

CD44 is a type I transmembrane glycoprotein that mediates cell-cell and cell-matrix interaction through its affinity for hyaluronic acid (HA) and possibly through other parts of the extracellular matrix (ECM). CD44 is highly polymorphic, possesses a number of alternative splice variants and undergoes extensive post-translational modifications (1,2). Increased surface levels of CD44 are characteristic of T cell activation, and expression of the protein is upregulated during the inflammatory response. Research studies have shown that interactions between CD44 and HER2 are linked to an increase in ovarian carcinoma cell growth (1-3). CD44 interacts with ezrin, radixin and moesin (ERM), linking the actin cytoskeleton to the plasma membrane and the ECM (4-6). CD44 is constitutively phosphorylated at Ser325 in resting cells. Activation of PKC results in phosphorylation of Ser291, dephosphorylation of Ser325, disassociation of ezrin from CD44, and directional motility (4).

  1. Goodison, S. et al. (1999) Mol. Pathol. 52, 189-196.
  2. Cichy, J. and Puré, E. (2003) J. Cell Biol. 161, 839-843.
  3. Bourguignon, L.Y. et al. (1997) J. Biol. Chem. 272, 27913-27918.
  4. Legg, J.W. et al. (2002) Nat. Cell Biol. 4, 399-407.
  5. Yonemura, S. et al. (1998) J. Cell Biol. 140, 885-895.
  6. Tsukita, S. et al. (1994) J. Cell Biol. 126, 391-401.
Entrez-Gene Id
960
Swiss-Prot Acc.
P16070
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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Product # Size Price
53289S
100 µg $ 229.0