Overview

  • Product name
    Anti-CD34 antibody [QBEND-10]
    See all CD34 primary antibodies
  • Description
    Mouse monoclonal [QBEND-10] to CD34
  • Host species
    Mouse
  • Specificity
    This antibody recognizes the CD34 antigen, some cross reactions with vascular associated adventitia and some basement membranes. Ultrastructure studies, using an indirect immuno-gold technique on ultra thin sections of human umbilical artery, have revealed that this antigen is associated to a degree with the basoluminal surfaces of endothelium and shows a slight reaction with an intracytoplasmic vesicular compartment. Predominant staining is on the endothelial cell membrane. QBEND-10 immunoprecipitates a glycoprotein with a relative MW 110kD expressed on haematopoietic cells and on the established myeloid leukaemic cell line KG1A.
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human, Cynomolgus monkey, Rhesus monkey
    Does not react with: Rat, Sheep, Cow, Dog
  • Immunogen

    Other Immunogen Type corresponding to CD34. Human endothelial vesicles

  • Epitope
    This antibody's target epitope is distinct from those of other anti-CD34 reactive mabs as demonstrated by its lack of sensitivity to neuraminidase treatment, and fluorescence staining profiles during flow cytometry analysis of target cell populations.
  • Positive control
    • tonsil, bone marrow

Properties

Applications

Our Abpromise guarantee covers the use of ab8536 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
Flow Cyt 1/50.

Mmethanol fixed cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P 1/50 - 1/100. Do not perform antigen retrieval.
WB Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.

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

  • Overlay histogram showing Jurkat cells stained with ab8536 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8536, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in Jurkat cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Anti-CD34 antibody [QBEND-10] (ab8536) at 5 µg/ml + Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa
    Additional bands at: 100 kDa (possible glycosylated form)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labelling CD34 with ab8536 at 1µg/ml (1:1000). Blood vessels show positive staining.

    Detection system polymeric HRP. HIER with Pressure cooker pH6.

  • ab8536 staining CD34 in human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/2000 in blocking buffer) for 16 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labelling CD34 with ab8536 at 1µg/ml (1:1000). Blood vessels show positive staining.

    Detection system polymeric HRP. HIER with Pressure cooker pH6.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling CD34 with ab8536 at 1ug/ml.
    Detection system polymeric HRP.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human umbilical cord labeling CD34 with ab8536 at 1/1000 dilution. Tissue was fixed in formaldehyde and permeabilized with PBS. Heat mediated antigen retrieval was performed using a citrate pH 6 buffer. Tissue was blocked with 5% serum for 1 hour at room temperature. A biotinylated polyclonal goat anti-mouse secondary antibody was used at 1/100 dilution.

    See Abreview

  • ab8536 at 1/500 staining human umbilical cord tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with horse serum before incubation with the antibody for 16 hours. A biotinylated goat anti-mouse IgG was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Barbhuiya MA  et al. Identification of spleen tyrosine kinase as a potential therapeutic target for esophageal squamous cell carcinoma using reverse phase protein arrays. Oncotarget 9:18422-18434 (2018). Read more (PubMed: 29719615) »
  • Rozza-de-Menezes RE  et al. Prevalence and clinicopathological characteristics of lipomatous neurofibromas in neurofibromatosis 1: An investigation of 229 cutaneous neurofibromas and a systematic review of the literature. J Cutan Pathol 45:743-753 (2018). Read more (PubMed: 29959804) »
See all 26 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Question

Inquiry:
ANTIBODY DETAILS Antibody Code: AB8536 Batch Number: GR49632
GENERAL INFORMATION
Antibody Storage Conditions (temperature/reconstitution etc) 4C
Description of the Problem (high background, low signal, non-specific staining etc)
No positively stained structures despite being in a simultaneous batch of tissues being stained where the only variable was using the previously ordered same antibody and the new antibody
Sample (Species/Tissue/Cell Type/Cell Line etc)
Human liver tissue containing metastases from colorectal carcinoma
Fixation of Sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration)
Formalin fixed Paraffin embedded
Antigen Retrieval (Enzymatic method, Heat mediated technique etc)
No antigen retrieval
Permeabilization step N/A
Blocking Conditions (Buffer/time period, Blocking agent)
Endogenous Peroxidase Block (DAKO) 30min 20% Normal Goat Serum (Invitrogen) 20min
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
CD34 Monoclonal 1:250, 1hr at 40C then overnight at 4C
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, wash step)
HRP secondary anti-mouse Detection Method HRP/DAB detection (DAKO) Positive and negative controls used (please specify) Positively stained vessels were present on a simultaneous run of the same tissue using the previous order of the same antibody Negative controls were producing by using IgG (at same concentration) as a substitute for the primary antibody
OPTIMIZATION ATTEMPTS (PROBLEM SOLVING)
How many times have you tried the IHC? 4
Have you run a No Primary control? Yes
Do you obtain the same results every time? Yes What steps have you altered? Additional Notes Previously ordered the same antibody and on simultaneous runs with no positive staining with new antibody there was positive staining with the old antibody

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hearthe customer has had difficulty obtaining satisfactory results from this vial of theantibody.

The details you have kindly provided will provide us with vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably recieved a bad vial.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 23 2016

Answer

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab8536 has been added to your order number ######. This has Abcam order reference ####.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

Read More

Answer

Thank you for contacting us and sorry for the delay of our reply.

I have been informed that the datasheets have been corrected and updated.

ab49168,ab49515,ab49273, ab49759 : the unit sizes for these products are now inµL.

ab8536 is 50µg, this antibody is Protein A purified, not a tissue culture supernatant.

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

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Answer



I have asked my colleagues in charge of the datasheets for the correct information regarding all these tissue culture supernatant antibodies.

I will contact you again as soon as I receive the right information. Sorry for the inconvenience and thank you for your patience.

Read More

Question
Answer

Thank you for contacting us.

Here is the additional information for flow cytometry - in addition to the figure legend given on the datasheet:

Methanol is used at 80% to fix the cells.The cells are blocked for 30minutes and there is a washing step between addition of the primary and secondary.

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 patience while I contacted the lab.

The Exact epiotpe site for ab8536 has not been mapped, so we are unsure if the epitope is intra- or extracellular. However we do know that the predominant staining is on the endothelial cell membrane.

I am sorry that I could not help you further in this matter but please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting Abcam.

We have not tested ab8536 in flow cytometry in non-fixed cells, it may work in non-fixed cells but as we have not tested it, we cannot guarantee that it will work for you.

If you do test ab8536 in non-fixed cells, then we would greatly appreciate any feedback you may have concerning this (positive or negative).

I am sorry that I was not able to provide more help in this matter and I wish you success in your experiments.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting us. ab31630, can be used in IHC-P ab8536, can be used in IHC-P ab65865; IHC-P is yet not tested we are unable to confirm it. Can we suggest alternatives as ab34364, ab87951 and ab101500? ab33775; IHC-P is not a tested application we are unable to confirm its use. We suggest using alternative as ab846, ab51312 ab33854; IHC-P again is not a tested application for this ab. We can suggest using ab9496, ab59754 In case you are interested in buying ab65865, ab33775 and ab33854 only I can then offer you three Abreview discount codes which will give you next three antibodies free of charge. Please be advised that in some cases heat damages the antigenic epitope we therefore always recommend our customers optimizing the heat mediated antigen retrieval step. Let me know if you are interested in my offer; I will then send you the discount codes. The alternatives are recommended based on their use in IHC-P so I would also suggest carefully checking the datasheet particularly immunogen and species in case you plan to use these antibodies. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Umbilical cord)
Specification
Umbilical cord
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
Yes - PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Mr. Manoj Kumar Valluru

Verified customer

Submitted Dec 04 2009

1-10 of 19 Abreviews or Q&A

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