Recombinant

Recombinant Anti-CD63 antibody [KILL150A] (ab271286)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [KILL150A] to CD63
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-CD63 antibody [KILL150A]
    See all CD63 primary antibodies

  • Description

    Mouse monoclonal [KILL150A] to CD63

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Tissue, cells or virus. This information is considered to be commercially sensitive.

  • Positive control

    • WB: SK-MEL-28 and HUVEC whole cell lysate. Human lymph node tissue lysate. IHC-P: Human melanoma tissue. ICC/IF: SK-MEL-28 cells. Flow Cyt: SK-MEL-28 and NK-92 cells.

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

Applications

Our Abpromise guarantee covers the use of ab271286 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
WB Use a concentration of 0.978 µg/ml. Predicted molecular weight: 26 kDa.
IHC-P Use a concentration of 0.978 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use a concentration of 19.56 µg/ml.
ICC/IF Use a concentration of 19.56 µg/ml.

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
      • 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

    • Western blot - Anti-CD63 antibody [KILL150A] (ab271286)
      Western blot - Anti-CD63 antibody [KILL150A] (ab271286)
      All lanes : Anti-CD63 antibody [KILL150A] (ab271286) at 0.978 µg/ml

      Lane 1 : SK-MEL-28 (human malignant melanoma), whole cell lysate
      Lane 2 : HUVEC (human umbilical vein endothelial cell), whole cell lysate
      Lane 3 : Human lymph node tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

      Predicted band size: 26 kDa



      CD63 can undergo glycosylation as shown in lane 1, 2 and 3 (PMID: 9890706, 28740179).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1-2: 3 mins; Lane 3: 48 secs.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [KILL150A] (ab271286)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [KILL150A] (ab271286)

      Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling CD63 with ab271286 at 1/4000 dilution (0.978µg/ml) followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). Cytoplasmic staining on human melanoma. The section was incubated with ab271268 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [KILL150A] (ab271286)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD63 antibody [KILL150A] (ab271286)

      Immunohistochemical analysis of paraffin-embedded human breast tissue labeling CD63 with ab271286 at 1/4000 dilution (0.978µg/ml) followed by ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879). The section was incubated with ab271268 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Negative control: Nearly no staining on human breast.

      Secondary antibody only control: Secondary antibody is ready to use Goat Anti-mouse IgG H&L (HRP polymer) (ab214879).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [KILL150A] (ab271286)
      Immunocytochemistry/ Immunofluorescence - Anti-CD63 antibody [KILL150A] (ab271286)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-MEL-28 cells labelling CD63 with ab271286 at 1/50 dilution (19.56µg/ml), followed by ab150113 Goat Anti-mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing lysosome/endosomes (ab252919) co-staining in SK-MEL-28 cells.

      ab252919 Anti-CD63 rabbit monoclonal antibody was used to counterstain tubulin at 1/250 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Negative control 1: ab271286 at a 1/50 dilution (19.56µg/ml) followed by ab150080 at a 1/1000 dilution.

      Negative control 2: ab259919 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.

    • Flow Cytometry - Anti-CD63 antibody [KILL150A] (ab271286)
      Flow Cytometry - Anti-CD63 antibody [KILL150A] (ab271286)

      Flow cytometric analysis of NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell) cells labelling CD63 with ab271286 at 1/1000 dilution (19.56µg/ml) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

    • Flow Cytometry - Anti-CD63 antibody [KILL150A] (ab271286)
      Flow Cytometry - Anti-CD63 antibody [KILL150A] (ab271286)

      Flow cytometric analysis of SK-MEL-28 (human malignant melanoma) cells labelling CD63 with ab271286 at 1/1000 dilution (19.56µg/ml) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab271286? Please let us know so that we can cite the reference in this datasheet.

    ab271286 has not yet been referenced specifically in any publications.

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