Rat Anti-CD44 Recombinant Antibody (clone 9A4) (CAT#: HPAB-N0328-YC)

The antibody (9A4) is a rat monoclonal antibody to CD44, variant 6. This antibody reacts with mouse. The CD44, variant 6 Antibody (9A4) has been validated for the following applications: Western Blot, ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin and Immunohistochemistry-Frozen.

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WB

Figure 1 Expression of CD44v6 (9A4 antibody) and ICAM-1 (11C81 antibody) in 3LL cells or hepatocytes derived from Cd44+/+ (WT) or Cd44−/− (KO) mice measured by Western blot analysis.

Figure 1 Expression of CD44v6 (9A4 antibody) and ICAM-1 (11C81 antibody) in 3LL cells or hepatocytes derived from Cd44+/+ (WT) or Cd44−/− (KO) mice measured by Western blot analysis.

Olaku, V., Matzke, A., Mitchell, C., Hasenauer, S., Sakkaravarthi, A., Pace, G.,... & Orian-Rousseau, V. (2011). c-Met recruits ICAM-1 as a coreceptor to compensate for the loss of CD44 in Cd44 null mice. Molecular biology of the cell, 22(15), 2777-2786.

FC

Figure 2 HGF binding of HT29 cells or HepG2 cells measured by FC analysis.

Figure 2 HGF binding of HT29 cells or HepG2 cells measured by FC analysis.

Gray area, staining with secondary antibody (without HGF). Gray line, binding of HGF. Dotted line, binding of HGF in presence of ICAM-1 antibody. Black line, binding of HGF in presence of CD44v6 antibody.

Olaku, V., Matzke, A., Mitchell, C., Hasenauer, S., Sakkaravarthi, A., Pace, G.,... & Orian-Rousseau, V. (2011). c-Met recruits ICAM-1 as a coreceptor to compensate for the loss of CD44 in Cd44 null mice. Molecular biology of the cell, 22(15), 2777-2786.

WB

Figure 3 HGF-induced Erk phosphorylation in BSp73AS cells transiently transfected (Amaxa) with expression vectors for ICAM-1 or CD44v6 or an empty vector as indicated. Bottom, the expression of ICAM-1 and CD44v6 (1.1ASML antibody) monitored in Western blots.

Figure 3 HGF-induced Erk phosphorylation in BSp73AS cells transiently transfected (Amaxa) with expression vectors for ICAM-1 or CD44v6 or an empty vector as indicated. Bottom, the expression of ICAM-1 and CD44v6 (1.1ASML antibody) monitored in Western blots.

Olaku, V., Matzke, A., Mitchell, C., Hasenauer, S., Sakkaravarthi, A., Pace, G.,... & Orian-Rousseau, V. (2011). c-Met recruits ICAM-1 as a coreceptor to compensate for the loss of CD44 in Cd44 null mice. Molecular biology of the cell, 22(15), 2777-2786.

FC

Figure 4 CD44 expression on LB cells recovered from local growth, remote axillary lymph node 1 spleen and cell culture.

Figure 4 CD44 expression on LB cells recovered from local growth, remote axillary lymph node 1 spleen and cell culture.

Surface marker expression on LB cells directly isolated from lymphoid organs and local growth was examined by indirect immunofluorescence followed by flow cytometry. Left histogram: incubation with secondary antibody alone. Right histogram: incubation with specific and secondary antibodies. Similar results were obtained in 6 separate experiments.

Vogt Sionov and, R., & Naor, D. (1997). Hyaluronan‐independent lodgment of CD44+ lymphoma cells in lymphoid organs. International journal of cancer, 71(3), 462-469.

IHC

Figure 5 The expression of CD44 (IM7), CD44v6 (9A4) and smooth muscle actin (SMA) in prepubertal (2 week old) and pubertal (7 week old) mouse mammary glands.

Figure 5 The expression of CD44 (IM7), CD44v6 (9A4) and smooth muscle actin (SMA) in prepubertal (2 week old) and pubertal (7 week old) mouse mammary glands.

(A) Stained ducts from 2-week-old mice. (B) Stained ducts from 7-week-old mice. For each antibody, ducts are shown with no CD44 staining (upper duct in each left panel), CD44 staining restricted to the myoepithelium (central inset panel in each case) and CD44 staining in both myoepithelial and luminal epithelial layers (right panel in each case). (C) Stained ducts from 7-weekold mice showing CD44 expression in the terminal portions of mammary ducts. Transverse section through TEBs (upper right panel in each case) and terminal portions of ducts (upper left panel in each case) are shown. The lower panel for each antibody shows a longitudinal section through the terminal portion of a mammary duct. The arrows denote areas of ductal epithelium where CD44 expression can be seen in both the myoepithelial layer and the luminal layer. Note that while all of the myoepithelium is positive for CD44 expression, not all areas of the duct show CD44 expression in the luminal cells. Bar, 50 µm.

Hebbard, L., Steffen, A., Zawadzki, V., Fieber, C., Howells, N., Moll, J.,... & Sleeman, J. (2000). CD44 expression and regulation during mammary gland development and function. J Cell Sci, 113(14), 2619-2630.

IHC

Figure 6 The expression of CD44 and SMA in the mammary gland during pregnancy, lactation and involution. Positive staining is brown-red in color. The sections were counterstained with hematoxylin.

Figure 6 The expression of CD44 and SMA in the mammary gland during pregnancy, lactation and involution. Positive staining is brown-red in color. The sections were counterstained with hematoxylin.

(A) CD44v6 staining in early pregnancy. Left panel: SMA. Right panel: 9A4. (B) CD44 staining during lactation. Left panel: SMA. Central panel: 9A4 (left and central panels are serial sections from mammary glands 10 days into lactation). Right panel: IM7. (C) SMA and 9A4 staining of large collecting ducts 12 days into lactation (left and central panels, respectively). Right panel: CD44v6 expression during involution of the mammary gland 8 days after weaning. Bar, 50 µm.

Hebbard, L., Steffen, A., Zawadzki, V., Fieber, C., Howells, N., Moll, J.,... & Sleeman, J. (2000). CD44 expression and regulation during mammary gland development and function. J Cell Sci, 113(14), 2619-2630.


Specifications

  • Host Species
  • Rat
  • Type
  • Rat IgG1
  • Specificity
  • Human CD44
  • Species Reactivity
  • Human, Mouse
  • Clone
  • 9A4
  • Applications
  • WB, ELISA, IHC-P, IHC-Fr

Product Property

  • Purity
  • >95% as determined by SDS-PAGE and HPLC analysis
  • Buffer
  • PBS
  • Preservative
  • No preservatives
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Applications

  • Application Notes
  • Flow Cytometry
    A total of 1 × 10⁶ cells was incubated with human CD44v6 or ICAM-1 antibody (100 μg/ml) for 1 h at 37°C prior to induction with HGF. The cells were detached using 5 mM EDTA in PBS, centrifuged at 1200 rpm for 3 min, and washed three times with PBS. Cells were incubated with HGF antibody (5 μg/ml) in PBS for 1 h on ice, followed by three washes and incubation with FITC anti–mouse IgG (1 μg/ml) for 30 min on ice in the dark. After three washes the cells were resuspended in PBS and measured with a FACScan.

Target

  • Alternative Names
  • IN; LHR; MC56; MDU2; MDU3; MIC4; Pgp1; CDW44; CSPG8; HCELL; HUTCH-I; ECMR-III

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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