Key features and details
- Mouse monoclonal [MEM-263] to CD44
- Suitable for: IP, WB, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
製品名Anti-CD44 antibody [MEM-263]
CD44 一次抗体 製品一覧
製品の詳細Mouse monoclonal [MEM-263] to CD44
アプリケーション適用あり: IP, WB, Flow Cyt, IHC-Pmore details
エピトープextracellular (N-terminal) domain
- Flow Cyt: Human peripheral blood lymphocytes. IHC-P: Human Skin Melanoma. WB: HPB-ALL, HeLa and A549 cell lysates.
保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.097% Sodium azide
Concentration information loading...
精製度Protein A purified
特記事項（精製）Purified from TCS. Purity >95% by SDS-PAGE.
Our Abpromise guarantee covers the use of ab9524 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 2 µg/ml.|
|WB||Use a concentration of 2 µg/ml. Use under non reducing condition.|
|Flow Cyt||Use 0.1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
機能Receptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix interactions through its affinity for HA, and possibly also through its affinity for other ligands such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA plays an important role in cell migration, tumor growth and progression. Also involved in lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to numerous alternative splicing and post-translational modification events.
組織特異性Isoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells.
配列類似性Contains 1 Link domain.
ドメインThe lectin-like LINK domain is responsible for hyaluronan binding.
翻訳後修飾Proteolytically cleaved in the extracellular matrix by specific proteinases (possibly MMPs) in several cell lines and tumors.
O-glycosylated; contains more-or-less-sulfated chondroitin sulfate glycans, whose number may affect the accessibility of specific proteinases to their cleavage site(s).
Phosphorylated; activation of PKC results in the dephosphorylation of Ser-706 (constitutive phosphorylation site), and the phosphorylation of Ser-672.
- Information by UniProt
- LHR antibody
- BA-1 antibody
- CD 44 antibody
Human peripheral blood lymphocytes stained with ab9524 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab9524, 0.1μg/1x106 cells) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.
All lanes : Anti-CD44 antibody [MEM-263] (ab9524) at 2 µg/ml
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CD44 knockout HeLa cell lysate
Lane 3 : A549 cell lysate
Lane 4 : LnCAP cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 80 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab9524 observed at 80 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab9524 was shown to react with CD44 in wild-type HeLa cells in western blot. Loss of signal was observed when CD44 knockout sample was used. Wild-type and CD44 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab9524 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 2 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
IHC image of CD44 staining in Human Skin Melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9524, 0.5 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blotting of HPB-ALL cell lysate (non-reduced sample) stained by ab9524.Western blotting of HPB-ALL cell lysate (non-reduced sample) stained by ab9524.
ab9524 は 5 報の論文で使用されています。
- Fitzgerald HC et al. Idiopathic infertility in women is associated with distinct changes in proliferative phase uterine fluid proteins. Biol Reprod 98:752-764 (2018). PubMed: 29546322
- Zhang L et al. The extent of inflammatory infiltration in primary cancer tissues is associated with lymphomagenesis in immunodeficient mice. Sci Rep 5:9447 (2015). IHC-P . PubMed: 25819560
- Samy M et al. Loss of the malignant phenotype of human neuroblastoma cells by a catalytically inactive dominant-negative hTERT mutant. Mol Cancer Ther 11:2384-93 (2012). Human . PubMed: 22933702
- Morigi M et al. Alternative pathway activation of complement by shiga toxin promotes exuberant c3a formation that triggers microvascular thrombosis. J Immunol 187:172-80 (2011). Blocking ; Human . PubMed: 21642543
- Kim S et al. Alpha-synuclein induces migration of BV-2 microglial cells by up-regulation of CD44 and MT1-MMP. J Neurochem 109:1483-96 (2009). PubMed: 19457162