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Mouse/Rat/Porcine/Equine CD44 Antibody

Catalog #: AF6127
13 Citations 1 Review Datasheet / COA / SDS
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AF6127
AF6127-SP
Detection of Mouse and Rat CD44 by Western Blot.
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Product Details
Citations (13)
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Supplemental Products
Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Mouse/Rat/Porcine/Equine CD44 Antibody Summary

Species Reactivity
Mouse, Rat, Porcine, Equine
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.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant mouse CD44
Gln25-Thr224
Accession # NP_033981
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Flow Cytometry
2.5 µg/106 cells
See below
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.
 
Immunocytochemistry
5-15 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.

Scientific Data

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

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.

Flow Cytometry Detection of CD44 antibody in Mouse Splenocytes antibody by Flow Cytometry. View Larger

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).

Flow Cytometry Detection of CD44 antibody in Rat Splenocytes antibody by Flow Cytometry. View Larger

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).

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

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).

Flow Cytometry Detection of CD44 antibody in Equine PBMCs antibody by Flow Cytometry. View Larger

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).

Immunocytochemistry CD44 antibody in Mouse Splenocytes by Immunocytochemistry (ICC). View Larger

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.

Immunocytochemistry CD44 antibody in Porcine Mesenchymal Stem Cells by Immunocytochemistry (ICC). View Larger

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.

Simple Western Detection of Rat CD44 antibody by Simple Western<sup>TM</sup>. 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-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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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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.

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.
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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Citations for Mouse/Rat/Porcine/Equine CD44 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. CD44 contributes to hyaluronan-mediated insulin resistance in skeletal muscle of high fat-fed C57BL/6 mice
    Authors: Hasib A, Hennayake CK, Bracy DP et al.
    Am. J. Physiol. Endocrinol. Metab.
  2. Limiting extracellular matrix expansion in diet-induced obese mice reduces cardiac insulin resistance and prevents myocardial remodelling
    Authors: Musale, V;Murdoch, CE;Banah, AK;Hasib, A;Hennayake, CK;Dong, B;Lang, CC;Wasserman, DH;Kang, L;
    Molecular metabolism
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  3. Astrocytic CD44 Deficiency Reduces the Severity of Kainate-Induced Epilepsy
    Authors: Kruk, PK;Nader, K;Skupien-Jaroszek, A;Wójtowicz, T;Buszka, A;Olech-Kocha?czyk, G;Wilczynski, GM;Worch, R;Kalita, K;W?odarczyk, J;Dzwonek, J;
    Cells
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  4. Negative regulation of CD44st by miR-138-5p affects the invasive ability of breast cancer cells and patient prognosis after breast cancer surgery
    Authors: FX Jian, PX Bao, WF Li, YH Cui, HG Hong
    BMC Cancer, 2023-03-24;23(1):269.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Hyaluronan supports the limbal stem cell phenotype during ex vivo culture
    Authors: Sudan Puri, Isabel Y. Moreno, Mingxia Sun, Sudhir Verma, Xiao Lin, Tarsis F. Gesteira et al.
    Stem Cell Research & Therapy
  6. SYK-3BP2 pathway activity in parenchymal and myeloid cells is a key pathogenic factor in metabolic steatohepatitis
    Authors: C Luci, E Vieira, M Bourinet, D Rousseau, S Bonnafous, S Patouraux, L Lefevre, F Larbret, V Prod'homme, A Iannelli, A Tran, R Anty, B Bailly-Mai, M Deckert, P Gual
    Cellular and Molecular Gastroenterology and Hepatology, 2021-08-16;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  7. Deconstructed Microfluidic Bone Marrow On-A-Chip to Study Normal and Malignant Hemopoietic Cell-Niche Interactions
    Authors: Aleman J, George SK, Herberg S et al.
    Small
  8. Serglycin is involved in inflammatory response in articular mouse chondrocytes
    Authors: A D'Ascola, M Scuruchi, A Avenoso, G Bruschetta, S Campo, G Mandraffin, GM Campo
    Biochem. Biophys. Res. Commun., 2018-03-31;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  9. Deficient TSC1/TSC2-complex suppression of SOX9-osteopontin-AKT signalling cascade constrains tumour growth in tuberous sclerosis complex
    Authors: Fuquan Jin
    Hum. Mol. Genet, 2017-01-15;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  10. Immunogenomics reveal molecular circuits of diclofenac induced liver injury in mice
    Authors: Eun-Hee Lee, Jung-Hwa Oh, Saravanakumar Selvaraj, Se-Myo Park, Mi-Sun Choi, Reinhard Spanel et al.
    Oncotarget
  11. Uncovering the dual role of RHAMM as an HA receptor and a regulator of CD44 expression in RHAMM-expressing mesenchymal progenitor cells
    Authors: Mandana Veiseh, Sean J. Leith, Cornelia Tolg, Sallie S. Elhayek, S. Bahram Bahrami, Lisa Collis et al.
    Frontiers in Cell and Developmental Biology
  12. Recombinant human hyaluronidase PH20 does not stimulate an acute inflammatory response and inhibits lipopolysaccharide-induced neutrophil recruitment in the air pouch model of inflammation.
    Authors: Huang Z, Zhao C, Chen Y, Cowell J, Wei G, Kultti A, Huang L, Thompson C, Rosengren S, Frost G, Shepard H
    J Immunol, 2014-04-28;192(11):5285-95.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  13. Effect of exosomes from adipose-derived stem cells on the apoptosis of Schwann cells in peripheral nerve injury.
    Authors: Liu, C Y, Yin, G Et al.
    CNS Neurosci Ther

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Mouse/Rat/Porcine/Equine CD44 Antibody
By Anonymous on 09/27/2017
Application: Immunocytochemistry/Immunofluorescence Sample Tested: HUVEC human umbilical vein endothelial cells Species: Human

This antibody works great for my staining.

Immunocytochemistry/Immunofluorescence Mouse/Rat/Porcine/Equine CD44 Antibody AF6127