Rat CD44 Antibody

Catalog # Availability Size / Price Qty
AF6577
AF6577-SP
Rat CD44 Antibody in Western Blot
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Rat CD44 Antibody Summary

Species Reactivity
Rat
Specificity
Detects rat CD44 in direct ELISAs. Detects mouse and rat CD44 in Western blots. In direct ELISA, approximately 50% cross-reactivity with recombinant mouse is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant rat CD44
Gln22-Glu271
Accession # O70509
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Simple Western
10 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection of Mouse and Rat CD44 by Western Blot. View Larger

Detection of Mouse and Rat CD44 by Western Blot. Western blot shows lysates of rat brain tissue, rat lung tissue, NR8383 rat alveolar macrophage cell line, and mouse spleen tissue. PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Rat CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6577) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for CD44 at approximately 80-100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western Detection of Rat CD44 by Simple WesternTM. View Larger

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-Rat CD44 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6577) 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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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). Rat CD44 has a 21 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 ECD of CD44H, rat CD44 (aa 23‑271) shares 90%, 73%, 72%, 75% and 70% identity with corresponding mouse, 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.
Entrez Gene IDs
960 (Human); 12505 (Mouse); 25406 (Rat); 100126860 (Porcine)
Alternate Names
CD44 antigen; CD44 molecule (Indian blood group); CD44; CD44R; CDw44; cell surface glycoprotein CD44; chondroitin sulfate proteoglycan 8; CSPG8; ECMR-III; epican; Extracellular matrix receptor III; GP90 lymphocyte homing/adhesion receptor; HCAM; HCELL; hematopoietic cell E- and L-selectin ligand; Heparan sulfate proteoglycan; Hermes antigen; homing function and Indian blood group system; HUTCH-I; Hyaluronate receptor; IN; LHR; MC56; MDU2; MDU2CD44 antigen (homing function and Indian blood group system); MDU3; MDU3CDW44; MIC4; MIC4MGC10468; MUTCH-I; Pgp1; PGP-1; PGP-I; Phagocytic glycoprotein 1; Phagocytic glycoprotein I

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