Overview

  • Product name
    Anti-CD44 antibody [F10-44-2]
    See all CD44 primary antibodies
  • Description
    Mouse monoclonal [F10-44-2] to CD44
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Fr, IP, Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    CD44-positive cell preparation.

  • Positive control
    • In Flow Cytometry, this antibody gave a positive signal in peripheral blood lymphocytes. In IHC, this antibody gave a positive signal in human kidney carcinoma sections and in ICC this antibody gave a positive result in HeLa cell line.
  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab6124 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use a concentration of 0.5 - 2 µg/ml.
IP Use a concentration of 0.5 - 2 µg/ml.
Flow Cyt Use 0.5-1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 10 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 81 kDa.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    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.
  • Tissue specificity
    Isoform 10 (epithelial isoform) is expressed by cells of epithelium and highly expressed by carcinomas. Expression is repressed in neuroblastoma cells.
  • Sequence similarities
    Contains 1 Link domain.
  • Domain
    The lectin-like LINK domain is responsible for hyaluronan binding.
  • Post-translational
    modifications
    Proteolytically cleaved in the extracellular matrix by specific proteinases (possibly MMPs) in several cell lines and tumors.
    N-glycosylated.
    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.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • LHR antibody
    • BA-1 antibody
    • CD 44 antibody
    • CD44 antibody
    • CD44 antigen antibody
    • CD44 molecule (Indian blood group) antibody
    • CD44 molecule antibody
    • CD44_HUMAN antibody
    • CDw44 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
    • Hyaluronate receptor antibody
    • IN antibody
    • INLU-related p80 Glycoprotein antibody
    • MC56 antibody
    • MDU2 antibody
    • MDU3 antibody
    • MGC10468 antibody
    • MIC4 antibody
    • MUTCH1 antibody
    • PGP-1 antibody
    • PGP-I antibody
    • PGP1 antibody
    • Phagocytic glycoprotein 1 antibody
    • Phagocytic glycoprotein I antibody
    • Soluble CD44 antibody
    see all

Images

  • Immunohistochemical analysis of Human esophogeal keratinocytes, staining CD44 with ab6124.

    Keratinocytes were cultured in vivo to form an epithelium for 15 days before fixing in formaldehyde and embedding in paraffin. Primary antibody was incubated overnight at 4°C and secondary antibody for 30 minutes at 37°C. Staining was detected using DAB.
  • Overlay histogram showing peripheral blood lymphocytes stained with ab6124 (red line). The cells were incubated with the antibody (ab6124, 0.5µg/1x106 cells) for 30 min at 4ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/200 dilution for 30 min at 4ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed gating on peripheral blood lymphocytes.

  • ICC/IF image of ab6124 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6124, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • IHC image of CD44 staining in human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab6124, 5µg/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.

References

This product has been referenced in:
  • Hu B  et al. CD44 promotes cell proliferation in non-small cell lung cancer. Oncol Lett 15:5627-5633 (2018). Read more (PubMed: 29552200) »
  • Vdovikova S  et al. A Novel Role of Listeria monocytogenes Membrane Vesicles in Inhibition of Autophagy and Cell Death. Front Cell Infect Microbiol 7:154 (2017). Read more (PubMed: 28516064) »
See all 29 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde

Ahmar Aziz

Verified customer

Submitted Jul 08 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (BEL 7404)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
40 µg
Treatment
80 µM Doxorubicin for 8hrs/24hrs
Specification
BEL 7404
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. guoqing zhu

Verified customer

Submitted Jan 22 2016

Answer

Thank you for contacting Abcam.
Earlier today we discussed the Human Mesenchymal Stromal Cell Marker panel ab93758. Based on the information on the datasheet, I compiled the following information for you:
1. Positive selection (CD105, CD29, CD44, CD90)
2. Negative selection (CD45)
The stem cell panel ab93758 consists of 5 unconjugated primary antibodies. These antibodies can be used with the secondary antibodies against mouse or rabbit of your choice. You can therefore adapt the fluorochrome choice to the ability of your machine.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Answer

Thank you for contacting us.

Based on the information from the journal Jpn J. Cancer Res 89 283-90 March 1989 (Yutaka Tokue et. al.), it states that this monoclonal antibody can recognises CD44s. Although the experiment sample used in this study was breast tumour cells but I believe the molecule should be the same in liver cells.

I hope this information is helpful to you. Please do not hesitate to contact us should you need any further information.

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Answer

Thank you for contacting Abcam regarding ab6124.


This ab was affinity purified on a protein G column. Please let me know if this information is sufficient or if you require additional details.


I hope this information is helpful. Do not hesitate to contact me if you have any additional questions or concerns.

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Answer

Thank you for your enquiry and interest in our products.

I can confirm that this panel contains 5 primary antibodies which recognize human mesenchymal stromal cells: ab6124, ab10559, ab23894, ab44967, ab52971. The datasheets can be found at these sites:

https://www.abcam.com/cd44-antibody-f10-44-2-ab6124.html

https://www.abcam.com/cd45-antibody-ab10559.html

https://www.abcam.com/cd90--thy1-antibody-af-9-ab23894.html

https://www.abcam.com/cd105-antibody-105c02-ab44967.html

https://www.abcam.com/integrin-beta-1-antibody-ep1041y-carboxyterminal-end-ab52971.html

There are some very informative and useful websites summarizing the different CD markers. You may wish to recommend to your customer for further consideration:

https://docs.abcam.com/pdf/immunology/cdantigen_poster.pdf

http://pathologyoutlines.com/cdmarkers.html

If you need any further assistance in the future, please do not hesitate to contact me.

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Question
Answer

Thank you for contacting us.

The link actually does not show results, I am sorry for the inconvenience.

Alternatively you can type CD44 in the search box provided on the website. The system will automatically give you sorted anti CD44 antibodies. Please select the antibodies based on your requirements of clonality, immunogen, species and application.

The most common are ab119863, ab6124, ab51037, ab19622, ab65829, ab25340, ab24504, ab119348 and ab25579 etc.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

Unfortunately, we do not have an anti-CD44 antibody from the H90 clone in our catalogue. We do however have several other antibodies against this target which may be suitable for your customer. The catalogue number you mentioned (ab93574) does not match to an anti-CD44 antibody. The clone mouse monoclonal anti-CD44 clone F10-44-2 is available with many different conjugates which can be viewed from the following link:

https://www.abcam.com/index.html?pageconfig=searchresults&search=cd44&pt=1&sk=clonenum&sv=F10-44-2&sn=F10-44-2&l=3&fViewMore=1

The suitability of this clone will depend on the application and samples your customer intends to use. If you could give me a few more details about this I may be able to advise further as to whether this clone would be suitable or if one of our other anti-CD44 antibodies would be more appropriate.

I hope this information has been of some help. If you would like any further assistance please do let me know.

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Answer

Vielen Dank für die Anfrage. Dass ab46793 nicht auf der Liste auftaucht, bedeutet dass er nicht getestet wurde, ob er mit Maus kreuzreagiert. In der Tat ist ab46793 für die Immunhistochemie (IHC-P/Fr - neben anderen Anwendungen wie Durchflusszytometrie) garantiert. Allerdings ist dieser CD44- Antikörper mit dem Tandemkonjugat PE-Cy7 gekoppelt. Welche Anregungswellenlängen beziehungsweise Filter Du für die Detektion benötigst, kannst Du anhand des Spektrums ersehen (Details dazu in Spectra-Viewern von eBioscience http://www.ebioscience.com/resources/fluorplan-spectra-viewer.htm  oder Biolegend http://www.biolegend.com/spectraanalyzer etc.) und beim Hersteller des Mikroskops erfragen. Im Allgemeinen sind PE und Tandem-Fluorochrome wie PE-Cy7 jedoch recht instabil und besitzen eine schwache Fluoreszenz-Intensität, so dass sie in der Regel wenig für die Mikroskopie geeignet sind. Wie auf dem Datenblatt von ab46793 beschrieben, würde ich deswegen für die Detektion dieses CD44- Antikörpers in der IHC einen sekundären Antikörper wie zum Beispiel ab96879 oder ab98697 empfehlen (je nach Fluorochrom-Belegung bei einer Mehrfachfärbung beziehungsweise vorhandenem Filtersystem). Daher ist es eigentlich nicht notwendig, den Klon [F10-44-2] als PE-Cy7-Konjugat zu nehmen (es sei denn, es sind noch andere Experimente damit geplant), sondern Du kannst ihn auch unkonjugiert erwerben (ab6124): ab6124: Click here (or use the following: https://www.abcam.com/index.html?datasheet=6124). ab96879: Click here (or use the following: https://www.abcam.com/index.html?datasheet=96879). ab98697: Click here (or use the following: https://www.abcam.com/index.html?datasheet=98697). Ich hoffe, diese Informationen helfen Dir weiter. Bitte zögere nicht, Dich wieder bei uns zu melden, falls Du weitere Fragen hast. 

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Question

Here you are my answers to abcam in details regarding my project so please forward to them and hopfuly I can get their guide asap. I am isolating human hertwig's epithelial root sheath (HERS) cells from periodontal ligament (PDL). So the cells that I am isolating are epithelial that should express these two epithelial markers as published in the literature (E-cadherin and Pan-cytokeratin) and will compare them with Stem cells from Human exfoliated deciduous teeth (SHED) using another two anibodies (CD44) that should be highly expressed in SHED than HERS and (CD73) that will be expressed similarly in both cells. So for characterization I ordered these antibodies from abcam: 1-      Mouse monoclonal (HECD-1) to E Cadherin (ab1416) 2-      Mouse monoclonal (PCK-26) to pan Cytokeratin (ab6401) 3-      Mouse monoclonal (F10-44-2) to CD44 (ab6124) 4-      Mouse monoclonal (7G2) to CD73 (ab54217) -          Immunofluorescence staining steps (first time) HERS were cultured in 4 well culture slide and the second day we did the following: a-      Discard the medium b-     Wash 3 times with PBS each time two mins c-      Add 500 µL methanol to each well for 20 mins at 4°C d-     Discard methanol and wash 3 times with PBS each time two mins e-      Add 7 drops of blocking reagent (from Millipore/ comes in the kit of immunoperoxidase secondary detection system) keep it at room temperature for 1 hr. f-       Discard blocking reagent and add the primary antibodies (1:50 ratio) (each antibody was applied separately in each well of the same chamber slide) (we diluted the antibodies using PBS) g-      Keep the chamber slide in 4°C for overnight. h-     Then discard the antibodies, wash 3 times with PBS each time two mins i-        Add 500 µL of secondary antibody (anti mouse FITC) to each well and keep it for 1 hr at room temperature j-       Discard secondary antibody and wash 3 times with PBS each time two mins k-     Add 500 µL of DAPI to each well keep it for 1 hr in a dark room l-        Discard DAPI and wash properly with PBS m-   The cover of the chamber slide was chopped and fluorescence mounting medium was applied (from Dako: S302380) n-     Cover slip was placed and the observed under fluorescence microscope image analyzer Under microscope: We observed the following (as I sent you previously some images); Green cells with green background and blues nucleus. So how to get black background with only green cells and blue nucleus as we got used for immunofluoescence images? -          We did optimize the method (second time) using the same previous procedures but with primary antibodies 1:300 but we got the same result. -          We did optimize the method (third time) using the same previous steps but with these changes: a-      We used PBS with Tween-20 for proper washing b-     Apply methanol and keep it in -20°C for 5 mins c-      Primary antibodies were applied with ratio (1:500) and keep it for 9 hrs at 4°C d-     DAPI was applied for half hr in a dark room. -          But under microscope we got the same result. So we are planning to use primary antibodies 1:1000 and keep it for 3 hrs but we do really need your recommendation and help to get the correct result before we run the test. And waiting for your new quotation please as I did email you. Best regards,    

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Answer

Thank you for contacting us. The background should be black. I am sorry we never had a question like this before. It looks like this problem might be due to instrumentation then antibodies itself. I have following recommendations that might help; - Use less number of cells per well. - Check the viability and passage level of cells. - The cells must be healthy with low number of passages. - Avoid drying out plate between washings. - Check microscope settings. - Check if the blocking is sufficiently done. You can also use 3-5% BSA for 1-2 hour in PBST for blocking. - Use more diluted primary antibodies. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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1-10 of 15 Abreviews or Q&A

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