Overview

  • Product name
    Anti-CD3 epsilon antibody, prediluted
    See all CD3 epsilon primary antibodies
  • Description
    Rabbit polyclonal to CD3 epsilon, prediluted
  • Host species
    Rabbit
  • Specificity
    This antibody recognises CD3 epsilon.
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human, Monkey
    Predicted to work with: Mouse, Rat, Horse, Cat, Dog, Baboon
  • Immunogen

    Synthetic peptide corresponding to CD3 epsilon.

  • Positive control
    • Recombinant Human CD3 epsilon protein (ab114153) can be used as a positive control in WB. Human tonsil for Immunohistochemistry.

Properties

Applications

Our Abpromise guarantee covers the use of ab27434 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
IHC-P 1/1. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. In particular, boil tissue sections in 10mM citrate buffer pH 6.0 for 10 min and then cool at room temperature for 20 min.
WB Use at an assay dependent concentration. Predicted molecular weight: 19 kDa.

Target

  • Function
    The CD3 complex mediates signal transduction.
  • Sequence similarities
    Contains 1 Ig-like (immunoglobulin-like) domain.
    Contains 1 ITAM domain.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD3 epsilon antibody
    • CD3e antibody
    • CD3e antigen epsilon polypeptide (TiT3 complex) antibody
    • CD3E antigen epsilon polypeptide antibody
    • CD3E antigen, epsilon subunit antibody
    • CD3e molecule epsilon antibody
    • CD3e molecule, epsilon (CD3 TCR complex) antibody
    • CD3e molecule, epsilon (CD3-TCR complex) antibody
    • CD3E_HUMAN antibody
    • IMD18 antibody
    • T cell antigen receptor complex epsilon subunit of T3 antibody
    • T cell surface antigen T3/Leu 4 epsilon chain antibody
    • T cell surface glycoprotein CD3 epsilon chain antibody
    • T-cell surface antigen T3/Leu-4 epsilon chain antibody
    • T-cell surface glycoprotein CD3 epsilon chain antibody
    • T3E antibody
    • TCRE antibody
    see all

Images

  • Ab27434, prediluted, staining CD3 epsilon in paraffin embedded, formalin fixed human tonsil by Immunohistochemistry.
  • Anti-CD3 epsilon antibody, prediluted (ab27434) at 1/25 dilution + Jurkat cell lysate

    Predicted band size: 19 kDa
    Observed band size: 19 kDa



    This dilution in this image refers to the dilution used for a non-prediluted version of this product.

References

This product has been referenced in:
  • Song Y  et al. Inflammation, Endothelial Injury, and Persistent Pulmonary Hypertension in Heterozygous BMPR2-Mutant Mice. Am J Physiol Heart Circ Physiol : (2008). Read more (PubMed: 18552156) »
See 1 Publication for this product

Customer reviews and Q&As

Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 175529 DESCRIPTION OF THE PROBLEM This antibody binds non-specifically, we noted nuclear staining. SAMPLE Mouse' brain and spleen tissue. PRIMARY ANTIBODY Abcam rabbit-CD3 prediluted antibody abab27434. Already comes prediluted. Tissues were incubated for 10min or 30 min. We also tried 1 hr and overnight. DETECTION METHOD Used DAB and also flourescence (Alexa). POSITIVE AND NEGATIVE CONTROLS USED We used spleens as positive controls and found extensive non-specific binding. We also included some negative controls (no primary). ANTIBODY STORAGE CONDITIONS 4 degrees as recommended by the manufacturer. FIXATION OF SAMPLE Tissues were fixed in 4 % PFA. Incubated in methanol/H202 for 20 min prior to initiating washes in dH20 followed by PBS 1%. Tissues were paraffin embedded. ANTIGEN RETRIEVAL Tissues were heated to 99 degrees in either citrate buffer or EDTA. Tissues were allowed to cool down to room temperature before proceeding with washes. PERMEABILIZATION STEP No permeabilization step was used to protect surface antigens. BLOCKING CONDITIONS Used normal goat serum (5%) with PBS 1% to block and with primary and secondary antibody. We also tried BSA (3%) with NGS/ PBS. We blocked for 30 min or 1 hr before adding primary. SECONDARY ANTIBODY The secondary we used was labelled polymer HRP anti-rabbit. One dropp (100 ul) were added to each tissue and incubated for 1 hr. Tissues were washed with PBS 1% 3X for 5 minutes HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? many HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? We altered the time of incubation for the primary. We also tried EDTA instead of citrate buffer for antigen retrieval. We also tried ADDITIONAL NOTES We specifcally purchased this antibody because it should have worked on embedded tissue. We also find that it does not work on frozen sections as we tried that as well. At this point we've spent more time trying to resolve this issue and have not made any progress. We are extremely unhappy with your product and the handling of this matter.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. From your previous correspondence I can see that you have contacted us on several occasions in recent months. I am also sorry for the delay to your enquiry and a resolution to this matter. I have read your technical questionnaire and I have a few comments. You are largely applying the antibody in a manner that I would recommend. However, please can you detail the duration and pH of the antigen retrieval approach that you are using in addition to the effect of changing the antigen retrieval solution to EDTA; whether this improved or changed the nuclear or cytoplasmic distribution of the antibody staining. I thank you for your patience and look forward to your response.

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

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