Anti-CD44 Picoband® Antibody (monoclonal, 7H7)

Western blot analysis of CD44 using anti-CD44 antibody (M00052-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: rat C6 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-CD44 antigen affinity purified monoclonal antibody (Catalog # M00052-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for CD44 at approximately 82KD. The expected band size for CD44 is at 82KD.

(A) floating tumorspheres after 6 days of suspension culture. Scale bar = 100 μm. (B) CD44 + CD24 - expression in TM40D cell line.
Index in PubMed under a CC BY license. PMID: 18477410

CD44 + CD24 - expression under serum-free culture condition during serial passages.
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CD44 + CD24 - expression in cells treated with 0.35PPC of PE. (A) one week after chemotherapy. (B) two weeks after chemotherapy.
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Hoxc10 exists in mesodermal derived callus. The immunofluorescence of Sox9 and CD44 in the mandible homotopic grafting (A), femoral heterotopic grafting (B) and femoral homotopic grafting(C). Sox9 represents cartilage (red), CD44 is a BMSCs marker (green), and DAPI marks the nucleus. (Scale bars, 500 μm) (D–F) represent the immunofluorescence of CD105 in mandible homotopic grafting, femoral heterotopic grafting, and femoral homotopic grafting, respectively. The white dotted line shows the edge of the femoral graft and the mandibular graft, and the middle of the dotted line is the callus. (Scale bars, 200 μm) (G) Localization of Hoxc10 at callus in femoral heterotopic grafting. (Scale bars, 100 μm; Scale bars, 20 μm) The data are presented as the mean ± SD ( n = 6). * p < 0.05.
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The expression of CD44 and CD24 of histological sections by immunochemistry analysis. (A, B) CD44 and CD24 staining from the xenograft tumors driven by a single sorted cell, respectively. (C, D) CD44 and CD24 staining from the xenograft tumors driven by unsorted TM40D cells, respectively. (both 20 objective).
Index in PubMed under a CC BY license. PMID: 18477410

IF analysis of CD44 using anti-CD44 antibody (M00052-3).
CD44 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti-CD44 Antibody (M00052-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of U87 cells using anti-CD44 antibody (M00052-3).
Overlay histogram showing U87 cells stained with M00052-3 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with mouse anti- CD44 Antibody (M00052-3, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.