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    cd9-antibody-epr23105-121-ab236630.pdf

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Cardiovascular Blood Platelets
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD9 antibody [EPR23105-121] (ab236630)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (2)

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Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Immunoprecipitation - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Flow Cytometry - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)
  • Anti-CD9 antibody [EPR23105-121] (ab236630)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23105-121] to CD9
  • Suitable for: WB, ICC/IF, IHC-P, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Protein
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Recombinant Human CD9 protein (BSA and azide free) (ab174040)
Knockout
Product image
Human CD9 knockout HeLa cell line (ab255375)

View more associated products

Overview

  • Product name

    Anti-CD9 antibody [EPR23105-121]
    See all CD9 primary antibodies
  • Description

    Rabbit monoclonal [EPR23105-121] to CD9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, MCF7, A549, HCT 116, Human tonsil and Human colon lysates. IHC-P: Human spleen and Human bladder carcinoma tissues. ICC/IF: HCT 116 cells. Flow Cyt: 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23105-121
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Platelets
    • Cardiovascular
    • Blood
    • Other
    • Immunology
    • Cell Type Markers
    • CD
    • Platelets
    • Stem Cells
    • Embryonic Stem Cells
    • Surface Molecules
    • Developmental Biology
    • Reproduction
    • Fertilisation
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Intracellular

Associated products

  • Alternative Versions

    • Anti-CD9 antibody [EPR23105-121] - BSA and Azide free (ab263023)
    • Alexa Fluor® 488 Anti-CD9 antibody [EPR23105-121] (ab267502)
    • Alexa Fluor® 647 Anti-CD9 antibody [EPR23105-121] (ab267503)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD9 knockout HeLa cell line (ab255375)
  • KO cell lysates

    • Human CD9 knockout HeLa cell lysate (ab263754)
  • Recombinant Protein

    • Recombinant Human CD9 protein (BSA and azide free) (ab174040)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab236630 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 (2)
1/1000. Detects a band of approximately 22 kDa (predicted molecular weight: 25 kDa).

 

 

ICC/IF (1)
1/500.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

 

 

Flow Cyt
1/50.

 

 

IP
1/30.

 

 

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

 

 

ICC/IF
1/500.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

 

 

Flow Cyt
1/50.

 

 

IP
1/30.

 

 

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.
  • Target information above from: UniProt accession P21926 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 928 Human
    • Omim: 143030 Human
    • SwissProt: P21926 Human
    • Unigene: 114286 Human
    • 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 [EPR23105-121] (ab236630)
      Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
      All lanes : Anti-CD9 antibody [EPR23105-121] (ab236630) at 1/1000 dilution

      Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : CD9 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 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 25 kDa
      Observed band size: 18 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab236630 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

      ab236630 was shown to react with CD9 in wild-type HeLa cells in Western blot with loss of signal observed in CD9 knockout cell line ab255375 (CD9 knockout cell lysate ab263754). Wild-type HeLa and CD9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab236630 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD9 antibody [EPR23105-121] (ab236630)
      Immunocytochemistry/ Immunofluorescence - Anti-CD9 antibody [EPR23105-121] (ab236630)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labelling CD9 with ab236630 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HCT 116 cell line. Negative control: Raji (PMID: 8921952). ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    • Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
      Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
      All lanes : Anti-CD9 antibody [EPR23105-121] (ab236630) at 1/1000 dilution

      Lane 1 : Human tonsil lysate
      Lane 2 : 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 expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 3275469).  Exposure time: 3 minutes.

    • Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
      Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)
      All lanes : Anti-CD9 antibody [EPR23105-121] (ab236630) 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 This blot was developed using a higher sensitivity ECL substrate.  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 [EPR23105-121] (ab236630)
      Immunoprecipitation - Anti-CD9 antibody [EPR23105-121] (ab236630)

      CD9 was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab236630 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236630 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: ab236630 IP in HCT 116 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236630 in HCT 116 whole cell lysate.

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

      Exposure time: 30 seconds.

       

    • Flow Cytometry - Anti-CD9 antibody [EPR23105-121] (ab236630)
      Flow Cytometry - Anti-CD9 antibody [EPR23105-121] (ab236630)

      Flow cytometric analysis of Raji (human Burkitt's lymphoma B lymphocyte, Left) / HCT 116 (human colorectal carcinoma epithelial cell, Right) cells labelling CD9 with ab236630 at 1/50 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 [EPR23105-121] (ab236630)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)

      Immunohistochemical analysis of paraffin-embedded Human bladder carcinoma tissue labeling CD9 with ab236630 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human bladder carcinoma. The section was incubated with ab236630 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 [EPR23105-121] (ab236630)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)

      Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD9 with ab236630 at 1/100 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 ab236630 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.

    • Anti-CD9 antibody [EPR23105-121] (ab236630)
      Anti-CD9 antibody [EPR23105-121] (ab236630)

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (2)

    Publishing research using ab236630? Please let us know so that we can cite the reference in this datasheet.

    ab236630 has been referenced in 2 publications.

    • Wang S  et al. Transfer of microRNA-25 by colorectal cancer cell-derived extracellular vesicles facilitates colorectal cancer development and metastasis. Mol Ther Nucleic Acids 23:552-564 (2021). PubMed: 33510943
    • Mathieu M  et al. Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9. Nat Commun 12:4389 (2021). PubMed: 34282141

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-CD9 antibody [EPR23105-121]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - other (Purified exosomes from mice)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    5 µg
    Specification
    Purified exosomes from mice
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 01 2022

    Western blot abreview for Anti-CD9 antibody [EPR23105-121]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (human HELA cells and murine placental samples)
    Gel Running Conditions
    Reduced Denaturing (Non-reduced conditions were also trialled)
    Loading amount
    20 µg
    Specification
    human HELA cells and murine placental samples
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Antonia Hufnagel

    Verified customer

    Submitted Sep 20 2021

    Immunocytochemistry/ Immunofluorescence abreview for Anti-CD9 antibody [EPR23105-121]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Glioblastoma, brain)
    Permeabilization
    Yes - Saponin 0.5%
    Specification
    Glioblastoma, brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Maria C Johansson

    Verified customer

    Submitted Sep 07 2021

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