Anti-TCR alpha + TCR beta antibody [H57-597] (ab25336)

Submit a review Q&A (3)References (9)

Overview

  • Product name
    Anti-TCR alpha + TCR beta antibody [H57-597]
    See all TCR alpha + TCR beta primary antibodies
  • Description
    Armenian hamster monoclonal [H57-597] to TCR alpha + TCR beta
  • Host species
    Armenian hamster
  • Specificity
    ab25336 reacts with a common epitope of the ß chain of the mouse aß T cell receptor (TCR) complex.
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, IPmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    TCR affinity purified from T-cell hybridoma DO-11.10 (Mouse)

  • General notes

    ab25336 does not react with yd TCR-bearing cells. Plate-bound or soluble ab25336 activates aß TCR-bearing T cells. In vitro and in vivo, ab25336 can induce immature thymocytes to undergo apoptosis. In vivo administration of ab25336 has been shown to deplete aß TCR-bearing cells to near completion and prevent graft rejection.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituents: 0.87% Sodium chloride, 0.14% Monobasic dihydrogen sodium phosphate
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    Purified from tissue culture supernatant
  • Primary antibody notes
    ab25336 does not react with yd TCR-bearing cells. Plate-bound or soluble ab25336 activates aß TCR-bearing T cells. In vitro and in vivo, ab25336 can induce immature thymocytes to undergo apoptosis. In vivo administration of ab25336 has been shown to deplete aß TCR-bearing cells to near completion and prevent graft rejection.
  • Clonality
    Monoclonal
  • Clone number
    H57-597
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab25336 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 at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IF Use at an assay dependent concentration.

Target

  • Relevance
    The receptors on T cells consist of immunoglobulin like integral membrane glycoproteins containing 2 polypeptide subunits, alpha and beta, of similar molecular weight, 40 to 55 kD in the human. Like the immunoglobulins of the B cells, each T cell receptor subunit has, external to the cell membrane, an N terminal variable domain and a C terminal constant domain. T cell receptors recognise foreign antigens which have been processed as small peptides and bound to major histocompatibility complex molecules at the surface of antigen presenting cells. Each T cell receptor is a dimer consisting of one alpha and one beta chain or one delta and one gamma chain. In a single cell, the T cell receptor loci are rearranged and expressed in the order delta, gamma, beta, and alpha. If both delta and gamma rearrangements produce functional chains, the cell expresses delta and gamma. If not, the cell proceeds to rearrange the beta and alpha loci.
  • Cellular localization
    Type I membrane protein.
  • Database links
    • Alternative names
      • FLJ22602 antibody
      • MGC117436 antibody
      • MGC22624 antibody
      • MGC23964 antibody
      • MGC71411 antibody
      • T cell antigen receptor alpha polypeptide antibody
      • T cell antigen receptor beta polypeptide T cell receptor beta cluster antibody
      • T cell receptor alpha (V D J C) antibody
      • T cell receptor alpha chain C region antibody
      • T cell receptor alpha constant antibody
      • T cell receptor alpha locus antibody
      • T cell receptor beta cluster antibody
      • T cell receptor beta locus antibody
      • TCRA antibody
      • TCRB antibody
      • TCRD antibody
      • TRA antibody
      • TRAC antibody
      • TRB antibody
      see all

    References

    This product has been referenced in:
    • Liang W  et al. Core Fucosylation of the T Cell Receptor Is Required for T Cell Activation. Front Immunol 9:78 (2018). Read more (PubMed: 29434598) »
    • Shi YF  et al. In vivo administration of monoclonal antibodies to the CD3 T cell receptor complex induces cell death (apoptosis) in immature thymocytes. J Immunol 146:3340-6 (1991). Read more (PubMed: 1827483) »
    See all 9 Publications for this product

    Publishing research using ab25336? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    Dear xxxx,

    Thank you for your reply. We are very disappointed to hear that this Ab has not been tested in paraffin, since we already bought this antibody! Could you sent us the protocol for staining frozen sections, we will probably start with that.

    Best regards Joost van den Berg

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    Answer

    Thank you for your reply.

    I can understand your disappointment. I was under the impression that you had not purchased the antibody yet. Of course as you have we will honor the Abpromise guarantee. If you find that the antibody does not work in the staining of your paraffin embedded tissue sections we would provide you with a replacement antibody of your choice or a refund. I will help in whatever way I can to obtain the best staining.

    As I mentioned, the antibody has been used in staining of frozen sections. This was done on tissue from mouse spleen. The antibody ab25336 was incubated overnight with the tissue at 4°C. The sections were then washed and subsequently incubated with the secondary Abs for 3 h at room temperature. I would initially start with a 1/100 dilution of the antibody and optimize from there.

    We usually find antigen retrieval with citrate buffer (pH 6) to work well with most targets but it would probably be worthwhile to try Tris-EDTA pH 9 as well.

    I hope this information has been of some help. If I can be of any further assistance, please do not hesitate to let me know.

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    Question

    Phone call requesting a protocol for how the anti-TCR alpha + TCR beta antibody [H57-597] (ab25336) has been used in IHC-P, particularly the antigen retrieval used.

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    Answer

    Thank you for contacting us yesterday in regards to the anti-TCR alpha + TCR beta antibody [H57-597] (ab25336).

    As discussed over the phone, I have checked into how the antibody has been used in IHC. Unfortunately, I have found that the antibody has actually not been used in staining of paraffin embedded sections as you hope to use it but only in frozen sections to date. There was simply an error made in adding this application to the datasheet of the product. This will be updated and the customers of the product informed.

    Unfortunately this means I cannot give you protocol information relating to IHC-P. I am however able to offer you the Abtrial testing discount to try the antibody if you would be interested. This offer means that if you purchase ab25336 as normal, test the antibody in IHC-P and let us know of the results through an Abreview. No matter whether the results are positive or negative, you would then be eligible to claim the value of ab25336 off any product in our catalogue. More information on this offer can be found here:

    www.abcam.com/abtrial

    Please note that the antibody to be tested must be purchased, tested, the Abreview submitted and the free product claimed within a 4 month period.

    If you would be interested in participating in this scheme please do let me know as a discount code needs to be issued prior to the purchase of ab25336.

    I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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    Question

    Thank you for your response, I will answer the questions 1. Samples: What sort of samples, species, tissue or cell type you used? brain sections (50um) from C57BL6 mice 2. At what dilution of this primary antibody you applied? What buffer you used for diluting this product? We tried different concentrations of the ab (1:1000, 1:500 and 1:100) in Tris-buffered saline (TBS: 50mM Tris + 0.9% NaCl pH 7.4) containing 0.5% (v/v) Triton X-100 and 1% of Horse serum) 3. Fixation: What percentage of PFA, for how long and at what temperature you used for fixation? 4% PFA at room temperature during 48h. 4. Permeabilization: At what percentage of Triton you used? For how long? all the solutions, washes, blocking serum and primary ab incubations are in Tris-buffered saline (TBS: 50mM Tris + 0.9% NaCl pH 7.4) containing 0.5% (v/v) Triton X-100. The blocking with horse serum is for 1h and the incubation with the primary is for 48 h. (everything at room temperature). 5. Have you applied any positive control along with the samples? No.

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    Answer

    Thank you for taking the time to fill in our technical questionaire and send it in to us. I am sorry you have had difficulty obtaining satisfactory results in immunohistochemistry with this antibody. I have read through the details and would like to make a few suggestions that I hope will help if you have not tried them already. 1. Your incubation time with primary antibody is very long. I would suggest 1 hour at a 1:100 dilution or 4oC overnight. 2. The target protein (TCR) is a membrane protein. Triton 100 is not suitable for membrane proteins, as it is a very strong detergent and can disrupt the membrane proteins and therefore the epitope for the antibody. I would suggest trying Tween 20 instead. There should be no requirement for permeabilisation to enable the antibody to access the epitope. However, on some rare occasions, antibody epitopes of membrane proteins are on the cytoplasmic side of the membrane (this will obviously depend on the antibody clone). If you obtain no staining after changing to Tween 20, you could try an acetone permeabilisation step. 3. Have you tried enzymatic antigen retrieval with for example proteinase K or trypsin? Again, this may be more gentle. 4. To the best of my knowledge, the C57BL6 mice should express TCR. There should be no difference between this and the TCR receptor expressed by BALB mice. 5. I would strongly recommend including a positive control. This could be lymph node or spleen samples from mice. I hope this is helpful. Please do not hesitate to get back in touch if you require more assistance.

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