Overview

  • Product name
    Anti-Glycophorin A + B antibody [HIR2]
    See all Glycophorin A + B primary antibodies
  • Description
    Mouse monoclonal [HIR2] to Glycophorin A + B
  • Host species
    Mouse
  • Specificity
    The antibody recognizes N-terminal, homologous portion of glycophorins A (GPA) and B (GPB), (strongly to GPA, and weakly to GPB). The antibody is useful in erythroid cell development studies, because HIR2 antigen is expressed on early erythroblasts, late erythroblasts, erythroblasts, mature erythrocytes and the cells of erythroid cell lines K562 and HEL, but not on all other cells (mature erythrocytes are characteristically CD235a positive and CD45 and CD71 negative).
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Glycophorin A + B (N terminal).

  • Positive control
    • FACS: peripheral blood leukocytes

Properties

Applications

Our Abpromise guarantee covers the use of ab15009 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 Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. PubMed: 23286773
IP Use at an assay dependent concentration. PubMed: 23286773
ICC/IF Use at an assay dependent concentration. PubMed: 19083849

Target

  • Relevance
    Glycophorins A (GYPA) and B (GYPB) are major sialoglycoproteins of the human erythrocyte membrane which bear the antigenic determinants for the MN and Ss blood groups. GYPA gene consists of 7 exons and has 97% sequence homology with GYPB from the 5' UTR to the coding sequence encoding the first 45 amino acids. GYPB accounts for S, s and U specificities. GPA and GPB provide the cells with a large mucin-like surface and it has been suggested this provides a barrier to cell fusion, so minimizing aggregation between red blood cells in the circulation. In addition to the M or N and S or s antigens, that commonly occur in all populations, about 40 related variant phenotypes have been identified. These variants include all the variants of the Miltenberger complex and several isoforms of Sta; also, Dantu, Sat, He, Mg, and deletion variants Ena, S-s-U- and Mk. Most of the variants are resulted from gene recombinations between GYPA and GYPB. These antigens are expressed on early erythroblasts, late erythroblasts, erythroblasts, mature erythrocytes and the cells of erythroid cell lines K562 and HEL, but not on all other cells (mature erythrocytes are characteristically CD235a positive and CD45 and CD71 negative).
  • Cellular localization
    Type I membrane protein.
  • Database links
  • Alternative names
    • CD235a antibody
    • CD235b antibody
    • Erythroid lineage specific membrane sialoglycoprotein antibody
    • Glycophorin Erik antibody
    • Glycophorin HeP2 antibody
    • Glycophorin MiI antibody
    • Glycophorin MiIII antibody
    • Glycophorin MiV antibody
    • Glycophorin MiVI antibody
    • Glycophorin MiX antibody
    • Glycophorin SAT antibody
    • Glycophorin Sta type C antibody
    • GPA antibody
    • GPB antibody
    • GPB.NY antibody
    • GPErik antibody
    • GpMiIII antibody
    • GPSAT antibody
    • GYPA antibody
    • GYPB antibody
    • GYPHe.NY antibody
    • HGpMiIII antibody
    • HGpMiV antibody
    • HGpMiVI antibody
    • HGpMiX antibody
    • HGpMiXI antibody
    • HGpSta(C) antibody
    • Mi.V glycoprotein (24 AA) antibody
    • MN antibody
    • MN sialoglycoprotein antibody
    • MNS antibody
    • PAS 2 antibody
    • PAS 3 antibody
    • PAS2 antibody
    • PAS3 antibody
    • Sialoglycoprotein alpha antibody
    • Sialoglycoprotein delta antibody
    • SS active sialoglycoprotein antibody
    • SS antibody
    see all

Images

  • ab15009 staining Human normal lung tissue. Staining is localised to cellular membranes. Left panel: with primary antibody at 1 µg/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus, at RT. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins followed by blocking with Dako Protein block for 10 mins (containing casein 0.25% in PBS) then incubated with primary antibody for 20 mins and detected with Dako Envision Flex amplification kit for 30 mins. Colorimetric detection was completed with DAB for 5 mins. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • ICC/IF image of ab15009 staining of Glycophorin A + B in Human cord blood cells. The sections were incubated in 10% serum to block non-specific protein-protein interactions. The sections were then incubated with ab15009 (1:1000) for one hour at +24°C, followed by AlexaFluor®568 conjugated secondary antibody. Red staining of the human cord blood cell membrane was observed.

    See Abreview

  • Overlay histogram showing K562 cells stained with ab15009 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab15009, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Flow cytometry analysis of human peripheral blood cells labelling Glycophorin A + B with ab15009. A APC-conjugated goat anti-mouse IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Soderblom EJ  et al. Proteomic analysis of ERK1/2-mediated human sickle red blood cell membrane protein phosphorylation. Clin Proteomics 10:1 (2013). WB, IP ; Human . Read more (PubMed: 23286773) »
  • Zhang X  et al. Terminal differentiation and loss of tumorigenicity of human cancers via pluripotency-based reprogramming. Oncogene : (2012). Read more (PubMed: 22777357) »
See all 5 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

We do not have a CD71 antibody that has been tested in ELISA or any other kind of capture assay, but the monoclonal antibody ab116015 is specific for an extracellular epitope and can at least identify CD71 on cells. Capturing reticulocytes with the magnetic beads that you plan on conjugating to the antibody has not been attempted, as far as we know. Please note that the formulation of the antibody contains 1% BSA, which may interfere with the conjugation.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=116015).

We have a "clean-up" kit that will remove the BSA by purifying the antibody over a Protein A column, if you are interested.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=102784).

Please let me know if you have any questions.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (First trimester Human decidua)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization
No
Specification
First trimester Human decidua
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 03 2017

Question
Answer

Thank you for your patience. We have not determined the light chain type of ab15009 in house, but I have found this clone in the literature listed as being IgG2b kappa. You could therefore use ab18421 or ab18469 as an isotype control. Please let me know if you need any additional information!

Read More

Answer

Thank you for contacting us.

We have 4, FITC conjugated anti Glycophorin A antibodies.

https://www.abcam.com/Glycophorin-A-antibody-YTH89-1-FITC-ab28082.html
https://www.abcam.com/Glycophorin-A-antibody-FITC-ab112201.html
https://www.abcam.com/Glycophorin-A-antibody-HIR2-FITC-Aminoterminal-end-ab52452.html
https://www.abcam.com/Glycophorin-A-antibody-CLB-AME-1-FITC-ab51531.html

3 Phycoerythrin conjugated abs

https://www.abcam.com/Glycophorin-A-antibody-CLB-AME-1-Phycoerythrin-ab51632.html
https://www.abcam.com/Glycophorin-A-antibody-YTH89-1-Phycoerythrin-ab33387.html
https://www.abcam.com/Glycophorin-A-B-antibody-HIR2-Phycoerythrin-ab26016.html

The price for these antibodies for 1 vial would be

ab28082; £283 per 100 microgram
ab51632: £227 per 500 micro litre
ab112201; £227 per 100 microgram
ab33387: £227 50 tests
ab52452: £227 per 100 tests
ab51531; £227 per 100 tests
ab26016: £227 per 100 tests

ab26016 the concentration is 0.01mg/ml

All other information is present on the datasheets. The product will be dispatched same day for next day delivery.

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

Question
Answer

Thank you for contacting us.

Here is the paper I mentioned in our conversation. It discusses the technique you plan on using, magnetic bead isolation.

The isolation of reticulocyte-free human red blood cells. Exp Biol Med (Maywood). 2007 Dec;232(11):1470-6. PMID: 18040072

http://ebm.rsmjournals.com/content/232/11/1470.full

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 reply.

Unfortunately we only have one anti-Glycophorin A antibody where we also have the blocking peptide available. This is goat polyclonalab40844 (blocking peptide is catalogue number ab45821) which is raised against a peptide taken from the C-terminal end of human Glycophorin A. This is not in the extracellular domain of Glycophorin A.

We have not tested this antibody in ICC yet and to our knowledge, neither has any of our customers. This does not mean that it will not work, simply that we have not tested it as yet. As the epitope recognised by theantibody is cytoplasmic, some permiabilisationwill be required.

If you would be interested intryingab40844for yourICC experiment, you would be eligible forthe testing discount scheme that we run. This would involve you purchasingab40844, testingit in ICC, then letting us knowof the results (regardless of if they are positive or negative) through an Abreview. You would then be entitledto a free primary antibody of your choicefrom our catalogue.More information on this scheme can be found from the following link:

https://www.abcam.com/collaboratordiscount

If you would like to participatein this scheme please do let me know as a discount code needs to be issued prior to purchase of ab40844.

Alternatively, as discussedab15009 hasbeenshown tobe compatible with ICC staining. If you would prefer to use this and perform an isotypecontrol to determine the specificity ofthe staining, ab18428could be used as theisotype control.

I hope thisinformation has beenof help. If you require any furtherinformation please donot hesitate to contact us again.

Read More

Answer

Thank you for getting back to me.

If you want to perform ICCto detectGlycophorin Ain human cells I would suggest ab15009 would be a good product to use for this.As you know, we do not know specifically if it detect the extracellular domain or not, but it has been used in house to perform ICC staining ofhuman cord blood cells (as presented on the datasheet of the antibody) as well as by Thorogate et al.:

Thorogate R et al. A novel fluorescence-based method in forensic science for the detection of blood in situ. Forensic Sci Int Genet 2:363-71 (2008). PubMed: 19083849

It may be that to obtain optimum staining with your specific samples you may require some optimisation of the protocol, including the permiabilisation and fixation used but the antibody is covered by the Abpromise guarentee for use with human samples with ICC. This means that if you find that it does not work, you would be entitled to a replacement antibody, refund or a credit note. More information on the Abpromise can be found from the following link:

https://www.abcam.com/abpromise

I hope this informationhas been of help.If you have any further questions please do not hesitate to contact us again.

Read More

Answer

Thank you for providing that information. I am sorry to ask further questions but it will put me in a better position to offer you advice in which of the antibodies we have which may be the most appropriate for you.

Do you want to detect the extracellular domain because that is the domain you are studying or because you think this will be the easiest to perform the ICC protocol with? Is your aim to detect the extracellular specifically, or just theglycophorin A protein using ICC?

With this additional information I will hopefully be able to assess which antibodies from our catalogue may be most suitable for your experiment.

I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us and your interest in our products.

Unfortunately I am unable to provide you with any further information to that which is provided on the datasheet due to proprietary reasons. Additionally, we do not have the peptide available for sale.

If you wouldbe interested I could look intowhether we have asuitable alternative antibody where a blocking peptide is available. Inorder to do this could you please let me know what application andspecies you are looking to use with thisantibody.

I am sorry that I could not be of more help in this instance. If you have any further questions please do not hesitate to contact us again.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Cord blood)
Specification
Cord blood
Fixative
Methanol
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted May 25 2011

1-10 of 11 Abreviews or Q&A

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

Sign up