Recombinant Anti-CD9 antibody [EPR23105-121] (ab236630)
![Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-458930-anti-cd9-antibody-epr23105-121-western-blot-wildtype-hela-cd9-knockout-hela-a549-mcf7.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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).
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| ICC/IF | (1) |
1/500.
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| IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Flow Cyt |
1/50.
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| IP |
1/30.
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| 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.
|
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Flow Cyt
1/50.
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IP
1/30.
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Target
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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
modificationsProtein 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
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Database links
- Entrez Gene: 928 Human
- Omim: 143030 Human
- SwissProt: P21926 Human
- Unigene: 114286 Human
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Alternative names
- Tetraspanin 29 antibody
- 5H9 antibody
- 5H9 antigen antibody
see all
Images
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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.
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![Immunocytochemistry/ Immunofluorescence - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-3-anti-cd9-antibody-epr23105-121-immunocytochemistry-hct-116-human.jpg)
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.
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![Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-355080-anti-cd9-antibody-epr23105-121-western-blot-human.jpg)
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.
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![Western blot - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-355079-anti-cd9-antibody-epr23105-121-western-blot-hct-116-human.jpg)
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
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![Immunoprecipitation - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-5-anti-cd9-antibody-epr23105-121-immunoprecipitation-hct-116-human.jpg)
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.
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![Flow Cytometry - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-4-anti-cd9-antibody-epr23105-121-flowcytometry-hct-116-human.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-2-anti-cd9-antibody-epr23105-121-immunohistochemistry-bladder-carcinoma-human.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-1-anti-cd9-antibody-epr23105-121-immunohistochemistry-spleen-human.jpg)
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.
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![Anti-CD9 antibody [EPR23105-121] (ab236630)](https://www.abcam.com/ps/products/236/ab236630/Images/ab236630-8-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
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