Anti-CD44 antibody [MEM-263]

• 产品名称

  Anti-CD44抗体[MEM-263]
  参阅全部 CD44 一抗

• 描述

  小鼠单克隆抗体[MEM-263] to CD44

• 宿主

  Mouse

• 经测试应用

  适用于: WB, Flow Cyt, IHC-Pmore details

• 种属反应性

  与反应: Human
  

• 免疫原

  Tissue, cells or virus corresponding to African green monkey CD44. COS-7 cells
  Database link: A0A0D9QZF5

• 表位

  extracellular (N-terminal) domain

• 阳性对照

• Flow Cyt: Human peripheral blood lymphocytes. IHC-P: Human Skin Melanoma. WB: HPB-ALL, HeLa, MOLT-4 and A549 cell lysates.

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  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.

• 存储溶液

  pH: 7.40
  Preservative: 0.097% Sodium azide
  Constituent: PBS

• 纯度

  Protein A purified

• 纯化说明

  Purified from TCS. Purity >95% by SDS-PAGE.

• 克隆

  单克隆

• 克隆编号

  MEM-263

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab9524于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

• Entrez Gene: 960 Human

• Omim: 107269 Human

• SwissProt: P16070 Human

• Unigene: 502328 Human

• 别名

• CD44 antibody

• CD44 antigen antibody

• CD44 molecule (Indian blood group) antibody

• CD44 molecule antibody

• CD44_HUMAN antibody

• CDw44 antibody

• CDW44 antigen antibody

• Cell surface glycoprotein CD44 antibody

• chondroitin sulfate proteoglycan 8 antibody

• CSPG8 antibody

• ECMR-III antibody

• Epican antibody

• Extracellular matrix receptor III antibody

• GP90 lymphocyte homing/adhesion receptor antibody

• HCELL antibody

• hematopoietic cell E- and L-selectin ligand antibody

• Heparan sulfate proteoglycan antibody

• Hermes antigen antibody

• homing function and Indian blood group system antibody

• HSA antibody

• HUTCH-I antibody

• HUTCH1 antibody

• HUTCHI antibody

• Hyaluronate receptor antibody

• IN antibody

• INLU-related p80 Glycoprotein antibody

• MC56 antibody

• MDU2 antibody

• MDU3 antibody

• MGC10468 antibody

• MIC4 antibody

• MUTCH I antibody

• MUTCH1 antibody

• PGP-1 antibody

• PGP-I antibody

• PGP1 antibody

• Phagocytic glycoprotein 1 antibody

• Phagocytic glycoprotein I antibody

• Soluble CD44 antibody

• LHR antibody

• BA-1 antibody

• CD 44 antibody

• 

  Western blot - Anti-CD44 antibody [MEM-263] (ab9524)

  All lanes : Anti-CD44 antibody [MEM-263] (ab9524) at 2 µg/ml
  
  Lane 1 : MOLT-4 cells (human T lymphoblast; acute lymphoblastic leukemia)
  Lane 2 : HeLa cells (human epithelial cell line from cervix adenocarcinoma)
  
  Performed under non-reducing conditions.
  

• 

  Flow Cytometry - Anti-CD44 antibody [MEM-263] (ab9524)

  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.

• 

  Western blot - Anti-CD44 antibody [MEM-263] (ab9524)

  All lanes : Anti-CD44 antibody [MEM-263] (ab9524) at 2 µg/ml
  
  Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 2 : CD44 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
  Lane 4 : LNCaP (Human prostate cancer cell line) whole cell lysate
  
  Lysates/proteins at 20 µg per lane.
  
  Performed under reducing conditions.
  
  Observed band size: 80 kDawhy 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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD44 antibody [MEM-263] (ab9524)

  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 blot - Anti-CD44 antibody [MEM-263] (ab9524)

  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.