Overview

  • Product name
    Anti-CD63 antibody [NK1/C3]
    See all CD63 primary antibodies
  • Description
    Mouse monoclonal [NK1/C3] to CD63
  • Host species
    Mouse
  • Specificity
    This antibody is specific to a 53 kD protein, expressed on activated platelets. CD63 is a lysosomal membrane glycoprotein that is translocated to plasma membrane after platelet activation. It is also expressed on granulocytes, B cells and T cells.
  • Tested applications
    Suitable for: ICC, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Smooth plasma membrane fraction of MeWo cells.

  • General notes


    CD63 is expressed on activated platelets. CD63 is a lysosomal membrane glycoprotein that is translocated to plasma membrane after platelet activation. It is also present in monocytes and macrophages and is weakly expressed in granulocytes, B, and T cells. CD63 is identical to the melanoma-associated antigen ME491 and to the platelet antigen PTLGP40.

Properties

Applications

Our Abpromise guarantee covers the use of ab1318 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
ICC Use at an assay dependent concentration.
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt 1/100.

(Also see PMID 19129916).

 

 

 

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    This antigen is associated with early stages of melanoma tumor progression. May play a role in growth regulation.
  • Tissue specificity
    Dysplastic nevi, radial growth phase primary melanomas, hematopoietic cells, tissue macrophages.
  • Sequence similarities
    Belongs to the tetraspanin (TM4SF) family.
  • Cellular localization
    Cell membrane. Lysosome membrane. Late endosome membrane. Also found in Weibel-Palade bodies of endothelial cells. Located in platelet dense granules.
  • Information by UniProt
  • Database links
  • Alternative names
    • Lysosomal associated membrane protein 3 antibody
    • CD 63 antibody
    • CD63 antibody
    • CD63 antigen (melanoma 1 antigen) antibody
    • CD63 antigen antibody
    • CD63 antigen melanoma 1 antigen antibody
    • CD63 molecule antibody
    • CD63_HUMAN antibody
    • gp55 antibody
    • Granulophysin antibody
    • LAMP 3 antibody
    • LAMP-3 antibody
    • LAMP3 antibody
    • LIMP antibody
    • Lysosomal-associated membrane protein 3 antibody
    • Lysosome associated membrane glycoprotein 3 antibody
    • Mast cell antigen AD1 antibody
    • ME491 antibody
    • Melanoma 1 antigen antibody
    • Melanoma associated antigen ME491 antibody
    • Melanoma associated antigen MLA1 antibody
    • Melanoma-associated antigen ME491 antibody
    • MGC72893 antibody
    • MLA 1 antibody
    • MLA1 antibody
    • NGA antibody
    • Ocular melanoma associated antigen antibody
    • Ocular melanoma-associated antigen antibody
    • OMA81H antibody
    • PTLGP40 antibody
    • Tetraspanin 30 antibody
    • Tetraspanin-30 antibody
    • Tspan 30 antibody
    • Tspan-30 antibody
    • TSPAN30 antibody
    see all

Images

  • ab1318 staining human melanoma by IHC-P.
  • Overlay histogram showing HL60 cells stained with ab1318 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab1318, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-Mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HL60 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Aaberg-Jessen C  et al. Co-expression of TIMP-1 and its cell surface binding partner CD63 in glioblastomas. BMC Cancer 18:270 (2018). Read more (PubMed: 29523123) »
  • Khazen R  et al. Melanoma cell lysosome secretory burst neutralizes the CTL-mediated cytotoxicity at the lytic synapse. Nat Commun 7:10823 (2016). Read more (PubMed: 26940455) »
See all 11 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us. If you are not able to easily pellet the exosomes, I would indeed recommend to add 20 µl of the antibody solution to the exosomes in 100 µl PBS. You may need to increase the incubation time and or the volume of ab81436 added, in order to optimise the staining conditions for this particular setting. To our knowledge, this has never been done before, so I am sorry that we cannot give you more detailed information. However, I trust that you will succeed! Unfortunately, it is NOT possible to determine the EXACT concentration of ab81436: The concentration of the specific TAPA1 antibodies has not been determined before conjugation. Even if it had been, this concentration would be different from the concentration of the antibody after the conjugation procedure, as there is always a loss during the purification steps. Determining the protein concentration after the conjugation does not result in accurate measurements, because Phycoerythrin (PE) is a huge protein itself and thus would give false results, as my colleagues tried to explain earlier (in case you did not receive these emails, see below). We would therefore suggest to use the PE isotype control ab81200 in the same dilution than ab81436. For example, if you add 20 µl of ab81436 to 100 µl of your sample, than it is recommended to also use 20 µl of the isotype control ab81200 for 100 µl of sample. Also, I would like to draw your attention to the criteria to record information about the experimental overview and samples (including controls) set out by the flow cytometry community (e.g. http://www.flowcytometryuk.org/). These criteria are known as the Minimum Information about a Flow Cytometry Experiment (MIFlowCyt), and contain valuable information about isotype controls and FMO controls. I hope this information is helpful to you. Please do not hesitate to contact me if you need any further advice or information in this regard. Previously: I can confirm that the concentration of this antibody was not determined. Regrettably, it is not possible to correctly measure the antibody concentration of products with antibodies conjugated to proteins (such as PE). /If there are concentrations given for conjugated antibodies, it is usually the IgG concentration measured before the conjugation took place and therefore only an estimate./ Unfortunately, we do not currently have more information about the concentration of ab81436./ Antibody concentration is usually determined by measuring the protein concentration. This is not useful in this case since PE is also a protein.

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Answer

Thank you for contacting us with your question about these antibodies. We don't have exact concentrations for either of these products, but ab1318 is around 100 ug/mL. Since Ab92726 is provided as unpurified tissue culture supernatant, it is not possible to measure the concentration, but supernatant is estimated to have a concentration around 1-3 mg/mL of total protein. I hope this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (cancer)
Specification
cancer
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 04 2010

Answer

Thank you for your enquiry. The concentration is :100 ug/ml (micrograms per ml). Please do not hesitate to contact us if you need further information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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