CD44 Antibody

R&D Systems | Catalog # AF6127

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse, Rat, Porcine, Equine

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Blockade of Receptor-ligand Interaction, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all CD44 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant mouse CD44
Gln25-Thr224
Accession # NP_033981

Specificity

Detects mouse and rat CD44 in direct ELISAs and Western blots. In direct ELISAs, approximately 35% cross-reactivity with recombinant human CD44 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for CD44 Antibody

Detection of Mouse and Rat CD44 antibody by Western Blot.

Detection of Mouse and Rat CD44 by Western Blot.

Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line, mouse spleen tissue, mouse lymph node tissue, and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for CD44 at approximately 80 to 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of CD44 antibody in Mouse Splenocytes antibody by Flow Cytometry.

Detection of CD44 in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes were stained with Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
Detection of CD44 antibody in Rat Splenocytes antibody by Flow Cytometry.

Detection of CD44 in Rat Splenocytes by Flow Cytometry.

Rat splenocytes were stained with Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
Detection of CD44 antibody in Porcine Mesenchymal Stem Cells antibody by Flow Cytometry.

Detection of CD44 in Porcine Mesenchymal Stem Cells by Flow Cytometry.

Porcine mesenchymal stem cells were stained with Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126).
Detection of CD44 antibody in Equine PBMCs antibody by Flow Cytometry.

Detection of CD44 in Equine PBMCs by Flow Cytometry.

Equine peripheral blood mononuclear cells (PBMCs) were stained with Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126).
CD44 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

CD44 in Mouse Splenocytes.

CD44 was detected in immersion fixed mouse splenocytes using Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
CD44 antibody in Porcine Mesenchymal Stem Cells by Immunocytochemistry (ICC).

CD44 in Porcine Mesenchymal Stem Cells.

CD44 was detected in immersion fixed porcine mesenchymal stem cells using Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Detection of Rat CD44 antibody by Simple WesternTM.

Detection of Rat CD44 by Simple WesternTM.

Simple Western lane view shows lysates of NR8383 rat alveolar macrophage cell line, loaded at 0.2 mg/mL. A specific band was detected for CD44 at approximately 169 kDa (as indicated) using 10 µg/mL of Sheep Anti-Mouse/Rat/Porcine/Equine CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6127) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for CD44 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 1-6 µg/mL of this antibody will block 50% of the binding of 25 ng/mL biotinylated Hyaluronan (132 kDa) to immobilized Recombinant Mouse CD44 Fc Chimera (Catalog # 6127-CD) coated at 1 µg/mL (100 μL/well). At 20 µg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Mouse splenocytes, rat splenocytes, porcine mesenchymal stem cells, and equine peripheral blood mononuclear cells (PBMCs)

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes and porcine mesenchymal stem cells

Simple Western

10 µg/mL
Sample: NR8383 rat alveolar macrophage cell line

Western Blot

1 µg/mL
Sample: RAW 264.7 mouse monocyte/macrophage cell line, mouse spleen tissue, mouse lymph node tissue, and rat brain tissue

Reviewed Applications

Read 2 reviews rated 4.5 using AF6127 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD44

CD44 is a ubiquitously expressed protein that is the major receptor for hyaluronan and exerts control over cell growth and migration (1 - 5). Mouse CD44 has a 22 amino acid (aa) signal sequence, an extracellular domain (ECD) with a 100 aa hyaluronan-binding disulfide-stabilized link region and a 48-463 aa stem region, a 21 aa transmembrane domain, and a 72 aa cytoplasmic domain. Within the stem, ten variably spliced exons (v1-10, exons 6-15) produce multiple protein isoforms (1‑5). The standard or hematopoietic form, CD44H, does not include the variable segments (1‑5). Cancer aggressiveness and T cell activation have been correlated with expression of specific isoforms (2, 4). With variable N- and O-glycosylation and splicing within the stalk, CD44 can range from 80 to 200 kDa (1, 2). Within the N‑terminal invariant portion of the ECD (aa 23-222), mouse CD44 shares 92%, 77%, 77%, 79% and 71% identity with corresponding rat, human, equine, canine and bovine CD44, respectively. The many reported functions of CD44 fall within three categories (1, 2). First, CD44 binds hyaluronan and other ligands within the extracellular matrix and can function as a "platform" for growth factors and metalloproteinases. Second, CD44 is a co-receptor that modifies activity of receptors including MET and the ErbB family of tyrosine kinases. Third, the CD44 intracellular domain links the plasma membrane to the actin cytoskeleton via the ERM proteins, ezrin, radixin and moesin. CD44 can be synthesized in a soluble form (4) or may be cleaved at multiple sites by either membrane-type matrix metalloproteinases, or ADAM proteases to produce soluble ectodomains (6, 7). The cellular portion may then undergo gamma secretase-dependent intramembrane cleavage to form an A beta ‑like transmembrane portion and a cytoplasmic signaling portion that affects gene expression (8, 9). These cleavage events are thought to promote metastasis by enhancing tumor cell motility and growth (1, 2, 6).

References

  1. Pure, E. and R.K. Assoian (2009) Cell. Signal. 21:651.
  2. Ponta, H. et al. (2003) Nat. Rev. Mol. Cell Biol. 4:33.
  3. Screaton, G.R. et al. (1992) Proc. Natl. Acad. Sci. USA 89:12160.
  4. Lynch, K.W. (2004) Nat. Rev. Immunol. 4:931.
  5. Yu, Q. and B.P. Toole (1996) J. Biol. Chem. 271:20603.
  6. Nagano, O. and H. Saya (2004) Cancer Sci. 95:930.
  7. Nakamura, H. et al. (2004) Cancer Res. 64:876.
  8. Murakami, D. et al. (2003) Oncogene 22:1511.
  9. Lammich, S. et al. (2002) J. Biol. Chem. 277:44754.

Alternate Names

CD44, ECMR-III, HCAM, HCELL, LHR, MDU2, MDU3, MIC4, MUTCH-I, Pgp1

Entrez Gene IDs

960 (Human); 12505 (Mouse); 25406 (Rat); 100126860 (Porcine)

Gene Symbol

CD44

UniProt

NP_033981

Additional CD44 Products

Product Documents for CD44 Antibody

Certificate of Analysis

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Product Specific Notices for CD44 Antibody

For research use only

Customer Reviews for CD44 Antibody (2)

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Customer Images

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  • Mouse/Rat/Porcine/Equine CD44 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 09/27/2017
    This antibody works great for my staining.
    CD44 Antibody AF6127
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 23645665
    Species: Rat
    Verified Customer | Posted 01/08/2015

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