Recombinant
RabMAb

Recombinant Anti-CD9 antibody [EPR2949] (ab92726)

Overview

  • Product name
    Anti-CD9 antibody [EPR2949]
    See all CD9 primary antibodies
  • Description
    Rabbit monoclonal [EPR2949] to CD9
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human CD9 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: P21926

  • Positive control
    • WB: HeLa, HuT-78, MCF7 and U87-MG cell lysates. Mouse heart and kidney lysate; Rat brain lysate. IHC-P: Human papillary carcinoma , astrocytoma, brain, kidney and tonsil tissue; Rat spleen tissue. ICC/IF: Mouse peritoneal macrophages. IP: HeLa whole cell lysate (ab150035).
  • General notes

      

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92726 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 1/2000. Predicted molecular weight: 25 kDa.
IP 1/10 - 1/100.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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

  • All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/1000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell). Whole cell lysates with 5% NFDM/TBST
    Lane 2 : Daudi (Human Burkitt's lymphoma lymphoblast). Whole cell lysates. (negative/low expression control) with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

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


    Exposure time: 3 seconds
  • Immunohistochemical staining of paraffin embedded rat spleen with purified ab92726 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • ab92726 (purified) at 1/20 immunoprecipitating CD9 in 10 μg HeLa (Lanes 1 and 2, observed at 24 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST

    Dilution buffer and concentration: 5% NFDM/TBST

  • All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/10000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HuT-78 cell lysate
    Lane 3 : U87-MG cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 25 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab92726 showing positive staining in Normal kidney tissue.

  • All lanes : Anti-CD9 antibody [EPR2949] (ab92726) at 1/2000 dilution (purified)

    Lane 1 : mouse heart lysate
    Lane 2 : mouse kidney lysate
    Lane 3 : rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 25 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab92726 showing positive staining in Papillary carcinoma of thyroid gland tissue.

  • Unpurified ab92726 showing positive staining in Astrocytoma tissue.

  • Unpurified ab92726 showing positive staining in Normal brain tissue.

  • Immunohistochemical staining of paraffin embedded human tonsil with purified ab92726 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

References

This product has been referenced in:
  • Erwig MS  et al. Anillin facilitates septin assembly to prevent pathological outfoldings of central nervous system myelin. Elife 8:N/A (2019). Read more (PubMed: 30672734) »
  • Li R  et al. Exosome-mediated secretion of LOXL4 promotes hepatocellular carcinoma cell invasion and metastasis. Mol Cancer 18:18 (2019). Read more (PubMed: 30704479) »
See all 73 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Application
Western blot
Sample
Chicken Tissue lysate - whole (Chicken embryo (HH8))
Gel Running Conditions
Non-reduced Denaturing
Loading amount
50 µg
Specification
Chicken embryo (HH8)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 16 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin - Psoriasis)
Permeabilization
No
Specification
Skin - Psoriasis
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Acetone

Dr. Christian Ostalecki

Verified customer

Submitted Aug 07 2018

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Hep G2)
Specification
Hep G2
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 05 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (liver)
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Nov 07 2014

Application
Flow Cytometry
Fixation
Paraformaldehyde
Permeabilization
Yes - 70% methanol
Sample
Human Cell (HepG2)
Specification
HepG2
Gating Strategy
Live cells
Preparation
Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: Enzyme free

Abcam user community

Verified customer

Submitted Jun 20 2014

Application
Western blot
Loading amount
50000 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Human Cell lysate - whole cell (HepG2)
Specification
HepG2
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Apr 04 2014

Answer

We have not specifically tested this Anti-CD9 antibody [EPR2949] (ab92726) as a marker of urinary exosomes but I can confirm that it can be used in western blot on mouse, rat, and human samples. Additionally, I have done a Google search and found that CD9 is indeed an exosome marker therefore it can be used as you mentioned. For more details about CD9 please see the publication below.


Can urinary exosomes act as treatment response markers in prostate cancer? Journal of Translational Medicine 2009, 7:4

http://www.translational-medicine.com/content/7/1/4


If you are also looking for a second antibody and detection reagents to go with this ab92726, here are some recommendation.


https://www.abcam.co.jp/donkey-polyclonal-secondary-antibody-to-rabbit-igg-h-l-hrp-ab97064.html [1mg; 28,000 yen; WB dilution 1/5000 - 1/50000]


https://www.abcam.co.jp/ecl-western-blotting-substrate-kit-50-tests-ab65623.html

Read More

Answer

Thank you for contacting us.
Yes, this antibody should work in mouse for Immunofluorescence. The reference below (also on our datasheet) has used this antibody for IF in mouse.

Huang W et al. CD9 tetraspanin interacts with CD36 on the surface of macrophages: a possible regulatory influence on uptake of oxidized low density lipoprotein. PLoS One 6:e29092 (2011). WB, ICC/IF, IP ; Mouse . http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=22216174&dopt=Abstract





I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your inquiry.

I can confirm that the immunogen of the antibody ab92726 (Anti-CD9 antibody [EPR2949]) is located in the cytoplasmic part of the protein.

This antibody is tested and guaranteed for WB, IP, IHC-P, ICC and Mouse, Rat, Human. It was tested in flow cytometry and we found it not suitable for this application.

I hope this information is helpful and wish you good luck for your research.

Read More

Answer

Vielen Dank für Ihre Anfrage.

Leider haben wir keine Teströhrchen von unseren Produkten die wir versenden könnten. Wir garantieren alle Antikörper so zu funktionieren, wie wir sie beschreiben. Das heisst, fallssie denAntikörperab92726 auf Paraffinschnitten auf humanem oder murinem Gewebe benutzen, ist dieser von uns garantiert zu funktionieren. Sollte er dies nicht tun, werden wir ihn einen kostenlosen Ersatz oder eine Gutschrift verschicken, wie in unserer Garantie beschrieben. https://www.abcam.com/abpromise

Falls Sie den Antikörper für eine andere Anwendung/Technikbenutzen wollen, eine welche von uns nicht getestet und deshalb nicht garantiert ist,lassen Sie es uns bitte wissen. Wir werden dann schauen, ob wir Ihnen einen Testangebot machen könnten.

Ich weiss leider nicht, ob CD9 in all Ihren Geweben exprimiert ist. Ich weiss leider auch nicht, welche Gewebe und welcheStrukturen Sie genau anschauen möchten. Nach Swissprot (http://www.uniprot.org/uniprot/P21926) ist CD9 auf hematopoetischen Zellen und Epithelzellen.

Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Read More

1-10 of 14 Abreviews or Q&A

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