Recombinant
RabMAb

Recombinant Anti-CD9 antibody [EPR23105-125] (ab263019)

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Overview

  • Product name

    Anti-CD9 antibody [EPR23105-125]
    See all CD9 primary antibodies

  • Description

    Rabbit monoclonal [EPR23105-125] to CD9

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human CD9 aa 100-200. The exact sequence is proprietary.
    Database link: P21926

  • Positive control

    • WB: HCT 116, HEL, Human tonsil and Human colon lysates. IHC-P: Human spleen and cervical carcinoma tissues. FC: HCT 116 cells. IP: HCT 116 cell lysate.

  • General notes

    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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab263019 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
IP 1/30.

 

 
WB 1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 25 kDa).

 

 
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

 

 
Flow Cyt 1/500.

 

 
  • Application notes
    Is unsuitable for ICC/IF.

    • Target

    • Function

      Involved in platelet activation and aggregation. Regulates paranodal junction formation. Involved in cell adhesion, cell motility and tumor metastasis. Required for sperm-egg fusion.

    • Tissue specificity

      Expressed by a variety of hematopoietic and epithelial cells.

    • Sequence similarities

      Belongs to the tetraspanin (TM4SF) family.

    • Post-translational
      modifications

      Protein exists in three forms with molecular masses between 22 and 27 kDa, and is known to carry covalently linked fatty acids.

    • Cellular localization

      Membrane.
    • Information by UniProt

    • Database links

      • Alternative names

        • Tetraspanin 29 antibody
        • 5H9 antibody
        • 5H9 antigen antibody
        • Antigen defined by monoclonal antibody 602 29 antibody
        • Antigen defined by monoclonal antibody 60229 antibody
        • BA-2/p24 antigen antibody
        • BA2 antibody
        • BTCC 1 antibody
        • BTCC1 antibody
        • CD9 antibody
        • CD9 antigen antibody
        • CD9 antigen p24 antibody
        • CD9 molecule antibody
        • CD9_HUMAN antibody
        • Cell growth inhibiting gene 2 protein antibody
        • Cell growth-inhibiting gene 2 protein antibody
        • DRAP 27 antibody
        • DRAP27 antibody
        • GIG2 antibody
        • Growth inhibiting gene 2 protein antibody
        • Leukocyte antigen MIC3 antibody
        • MIC3 antibody
        • Motility related protein antibody
        • Motility-related protein antibody
        • MRP 1 antibody
        • MRP-1 antibody
        • MRP1 antibody
        • p24 antibody
        • p24 antigen antibody
        • Tetraspanin-29 antibody
        • Tspan 29 antibody
        • Tspan-29 antibody
        • TSPAN29 antibody
        see all

      Images

      • Western blot - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Western blot - Anti-CD9 antibody [EPR23105-125] (ab263019)
        All lanes : Anti-CD9 antibody [EPR23105-125] (ab263019) at 1/1000 dilution

        Lane 1 : HEL (erythroleukemia erythroblast), whole cell lysate
        Lane 2 : Human tonsil lysate
        Lane 3 : Human colon lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

        Predicted band size: 25 kDa
        Observed band size: 22 kDa
        why is the actual band size different from the predicted?



        Blocking and diluting buffer and concentration: 5% NFDM/TBST.

        The molecular weight observed is consistent with what has been described in the literature (PMID: 3275469).  Exposure time: 3 minutes.

      • Western blot - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Western blot - Anti-CD9 antibody [EPR23105-125] (ab263019)
        All lanes : Anti-CD9 antibody [EPR23105-125] (ab263019) at 1/1000 dilution

        Lane 1 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate
        Lane 2 : Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

        Predicted band size: 25 kDa
        Observed band size: 22 kDa why is the actual band size different from the predicted?



        Blocking and diluting buffer and concentration: 5% NFDM/TBST.

        The molecular weight observed is consistent with what has been described in the literature (PMID: 3275469). 

        Negative control: Raji (PMID: 8921952). Exposure time: 3 minutes.

      • Immunoprecipitation - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Immunoprecipitation - Anti-CD9 antibody [EPR23105-125] (ab263019)

        CD9 was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate  with ab263019 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab263019 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

        Lane 1: HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate 10ug.

        Lane 2: ab263019 IP in HCT 116 whole cell lysate.

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab263019 in HCT 116 whole cell lysate.

        Blocking and dilution buffer and concentration: 5% NFDM/TBST.

        Exposure time: 10 seconds.

         

      • Flow Cytometry - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Flow Cytometry - Anti-CD9 antibody [EPR23105-125] (ab263019)

        Flow cytometric analysis of Raji (human Burkitt's lymphoma B lymphocyte, Left) / HCT 116 (human colorectal carcinoma epithelial cell, Right) cells labelling CD9 with ab263019 at 1/500 dilution 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 was used as the secondary antibody.

        Negative control: Raji (PMID: 8921952).

        Gated on viable cells.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-125] (ab263019)

        Immunohistochemical analysis of paraffin-embedded Human cervical carcinoma tissue labeling CD9 with ab263019 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cervical carcinoma. The section was incubated with ab263019 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

        Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-125] (ab263019)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-125] (ab263019)

        Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD9 with ab263019 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on platelets of human spleen. The section was incubated with ab263019 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 a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

        Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      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

      Publishing research using ab263019? Please let us know so that we can cite the reference in this datasheet.

      ab263019 has not yet been referenced specifically in any publications.

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