Overview

  • Product name
    Anti-CD34 antibody [EP373Y]
    See all CD34 primary antibodies
  • Description
    Rabbit monoclonal [EP373Y] to CD34
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IP, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Dog, Human, African bush elephant
  • Immunogen

    Synthetic peptide within Human CD34 aa 350 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P28906

  • Positive control
    • Western Blot control TF-1. IHC controls Human kidney, HUVEC cells and rat kidney. This antibody gave a positive result in IHC in the following FFPE tissue: Mouse normal brain. IHC-P: Mouse prostate and kidney tissues. IHC-Fr: Mouse lung tissue.
  • 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 ab81289 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 1/10000. Detects a band of approximately 120 kDa (predicted molecular weight: 41 kDa).

For unpurified use at 1/10000 - 1/100000.

ab81289 is not suitable for rat and mouse species in WB application. 

IHC-P 1/2500 - 1/5000.

See IHC antigen retrieval protocols.

For unpurified use at  1/100 - 1/250.

ICC/IF 1/100 - 1/250.

 

IP 1/30.
IHC-Fr 1/100.
Flow Cyt Use at an assay dependent concentration.

Target

  • Function
    Possible adhesion molecule with a role in early hematopoiesis by mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Could act as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components. Presents carbohydrate ligands to selectins.
  • Tissue specificity
    Selectively expressed on hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues.
  • Sequence similarities
    Belongs to the CD34 family.
  • Developmental stage
    On early hematopoietic progenitor cells.
  • Post-translational
    modifications
    Highly glycosylated.
    Phosphorylated on serine residues by PKC.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD34 antibody
    • CD34 antigen antibody
    • CD34 molecule antibody
    • CD34_HUMAN antibody
    • Cluster designation 34 antibody
    • Hematopoietic progenitor cell antigen CD34 antibody
    • HPCA1 antibody
    • Mucosialin antibody
    • OTTHUMP00000034733 antibody
    • OTTHUMP00000034734 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling CD34 with purified ab81289 at 1/2500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Negative control using PBS instead of primary antibody. Counter stained with Hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of HUVEC (Human umbilical vein endothelial cell line) cells labelling CD34 with purified ab81289 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain.

    Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Flow Cytometry analysis of TF-1(human erythroleukemia ) cells labeling CD34 with purified ab81289 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • purified at 1/10000 dilution + TF-1 (Human bone marrow erythroleukemia cells) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 41 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?



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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human angiosarcoma labeling CD34 with unpurified ab81289 at 1/100-1/250.

  • Paraformaldehyde-fixed, paraffin-embedded mouse prostate tissue stained for CD34 using ab81289 at 1/150 dilution in immunohistochemical analysis.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling CD34 with purified ab81289 at 1/2500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody at 1/500. Negative control using PBS instead of primary antibody. Counter stained with Hematoxylin.

  • Frozen mouse lung tissue stained for CD34 using ab81289 at 1/400 dilution in immunohistochemical analysis, followed by donkey anti-rabbit Alexa Fluor® 546 at 1/2000 dilution.

    See Abreview

  • Paraformaldehyde-fixed, paraffin-embedded mouse kidney tissue stained for CD34 using ab81289 at 1/200 dilution in immunohistochemical analysis.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal tonsil vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.

     

     

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • CD34 was immunoprecipitated from TF-1 (Human bone marrow erythroleukemia cells) cells using purified ab81289 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab81289. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution. Blocking and dilution buffer and concentration: 5% NFDM/TBST

  • Immunocytochemistry/Immunofluorescence analysis of human embryonic stem cell-derived endothelial cells labeling CD34 with unpurified ab81289 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% aTriton X-100. An Alexa Fluor® 647-conjugated secondary antibody was used at a 1/400 dilution. DAPI (blue) was used as the nuclear counter stain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal kidney vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Normal uterus vessels tissue labeling CD34 with unpurified ab81289 at 1/100-1/250.


     

  • IHC image of CD34 staining in Mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab81289, 1/250 dilution, for 15 mins at room temperature. A Goat anti-Rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:
  • Tang YC  et al. Ginsenoside Rg3 targets cancer stem cells and tumor angiogenesis to inhibit colorectal cancer progression in vivo. Int J Oncol 52:127-138 (2018). Read more (PubMed: 29115601) »
  • Mizoguchi F  et al. Functionally distinct disease-associated fibroblast subsets in rheumatoid arthritis. Nat Commun 9:789 (2018). ICC/IF . Read more (PubMed: 29476097) »
See all 131 Publications for this product

Customer reviews and Q&As

1-10 of 15 Q&A

Answer

Thank you for your patience while I looked into this for you.
Rabbits do not have specific IgG subclasses like mice or humans. Therefore, all of our rabbit monoclonals are simply considered to be rabbit IgG.
I hope this helps. Please let me know if you have any additional questions.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number Order 1205432.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question
Answer

Thank you for contacting us.

I have received an answer from laboratory. unfortunately they have confirmed that the antibody wasn't tested with sheep samples so we will not be able to guarantee this. This antibody was indeed tested positively in flow cytometry using human cell samples using the protcol given in the link http://www.epitomics.com/pdf/FCP1.pdf.

On further investigation about the sheep species I realized that sheep has been added because a received abreviw where one of our customer have used this antibody on sheep tissue section sin IHC-P application (https://www.abcam.com/index.html?datasheet=81289&tab=abreviews&intabreviewid=21740).

I would suggest using human cells as positive control and checking, if the problem is due to antibody or protocol.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

leider muss ich Innen bestätigen, dass wir keine perfekte Kombination von Antikörpern haben, für Ihre Versuche.

Wenn ich Sie richtig verstanden habe, möchten Sie einen IF auf gefrorenem Gewebe machen?

ab81289 (Anti-CD34 antibody [EP373Y]) ist getestet für IHC-P und ICC/IF und funktioniert deshalb mit höchster Wahrscheinlichkeit auf für IHC-Fr.

Bei einer Mehrfachfärbung wie dieser, kann ich außerdem empfehlen pre-absorbierte sekundäre Antikörper zu verwenden.


ab30446 (Anti-CD45 antibody [YTH24.5]) ist nur für Durchflusszytometrie getestet.


Unseres Wissens nach wurde ab30446 bisher noch nicht in IHC getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab30446 abdecken, und gegen eine erneute Bestellung weiterer Produkte von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab30446 in IHC testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in IHC.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können weitere Produkte zu dem gleichen Preis wie ab30446bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

Read More

Answer

Hello! The datasheets have been updated and any "SDS" necessary should be available for pdf download from our online datasheet. Please let me know if you have any further questions.

Read More

Answer

Thank you for contacting Abcam. For future reference, all requests for MSDS should go to mailto:product.updates@abcam.com. I have copied them on this email and they should get back to you shortly!

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for contacting us.
You are right, the lab tested this antibody in Flow Cytometry on Human U937 cells, not on pig cells.
The cross reactivity for pig cells was added to the product datasheet based on a result submitted by one of our customers. The results have been published on our website via our Abreview system. The link to the data and protocol is https://www.abcam.com/index.html?datasheet=81289&tab=abreviews&intabreviewid=26886
I am sorry that this antibody did not perform in Pig samples as stated on the datasheet, I have asked our Finance department to issue a credit note for you.
Your credit note ID is 21736.
The credit note may be used in one of the following ways:
(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.
Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.
To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.
The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Answer

Thank you for your reply.
I received the following information from the lab:
ab81289 was tested in Flow Cytometry on permeabilized U937 cells. Unfortunately, we do not have a dot blot data, but attached is the FC data for this antibody.
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 taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab81289. I would also appreciate if you can confirm some further details:

- What is the lot number of ab81289 you received and tested?
- This antibody was tested in flow cytometry on Pig Cell (Blood buffy coat).The cells were fixedusing methanol, not permeabilised, the antibody was incubated for 30 minutesat 4ºC in the presence of 3% BSA.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More

1-10 of 15 Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up