Overview

  • Product name
  • Description
    Rabbit polyclonal to ECSIT
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to 14 amino acids near the C-terminus of ECSIT (Human).

  • Positive control
    • human heart lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab21288 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
ICC/IF Use at an assay dependent concentration. PubMed: 21163940
IP Use at an assay dependent concentration. PubMed: 21163940
IHC-P Use a concentration of 2 µg/ml.
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 49 kDa).

Target

  • Function
    Adapter protein of the Toll-like and IL-1 receptor signaling pathway that is involved in the activation of NF-kappa-B via MAP3K1. Promotes proteolytic activation of MAP3K1. Involved in the BMP signaling pathway. Required for normal embryonic development (By similarity). Required for efficient assembly of mitochondrial NADH:ubiquinone oxidoreductase.
  • Sequence similarities
    Belongs to the ECSIT family.
  • Cellular localization
    Cytoplasm. Nucleus. Mitochondrion.
  • Information by UniProt
  • Database links
  • Alternative names
    • Ecsit antibody
    • ECSIT homolog antibody
    • ECSIT protein antibody
    • ECSIT_HUMAN antibody
    • Evolutionarily conserved signaling intermediate in Toll pathway, mitochondrial antibody
    • Evolutionarily conserved signaling intermediate in Toll pathways antibody
    • Likely ortholog of mouse signaling intermediate in Toll pathway evolutionarily conserved antibody
    • Protein SITPEC antibody
    • Signaling intermediate in Toll pathway evolutionarily conserved antibody
    • SITPEC antibody
    • SITPEC protein antibody
    see all

Images

  • Lane 1 : Anti-ECSIT antibody (ab21288) at 0.5 µg/ml
    Lane 2 : Anti-ECSIT antibody (ab21288) at 1 µg/ml
    Lane 3 : Anti-ECSIT antibody (ab21288) at 2 µg/ml

    All lanes : human heart lysate

    Predicted band size: 49 kDa
    Observed band size: 42 kDa
    why is the actual band size different from the predicted?

  • ab21288 at 2µg/ml staining ECSIT in mouse heart by IHC

References

This product has been referenced in:
  • Guarani V  et al. TIMMDC1/C3orf1 functions as a membrane-embedded mitochondrial complex I assembly factor through association with the MCIA complex. Mol Cell Biol 34:847-61 (2014). Read more (PubMed: 24344204) »
  • Soler-López M  et al. Interactome mapping suggests new mechanistic details underlying Alzheimer's disease. Genome Res 21:364-76 (2011). WB, ICC/IF, IP ; Human . Read more (PubMed: 21163940) »
See all 3 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your enquiry and your interest.

I regret to inform you that the immunogen sequence is commercially sensitive information and we are not allowed to release it. However, it is between residues 301 and 350 aa accession number NP_057665.

Since antibody is a polyclonal antibody, it recognizes different epitopes so epitope mapping is not performed.

If you need any further assistance, please do not hesitate to contact me.

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Answer

Thank you very much for your reply and Nezira's. At this stage I think it would be worth looking at Nezira's protocol in detail in light of the fact that the first lot did not give her the results expected. Can you please ask Nezira to click on the link below and fill in our online questionnaire for WB, we will then be able to review all her data and understand the source of the problem. I would already recommend to run a positive control of human heart lysate to make sure that the problem is not due to her sample preparation. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=21288&mode=questionaire We look forward to receiving this information to resolve this matter, thank you both in advance,

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Question

BATCH NUMBER 130274 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Wrong band. It detects a band that looks like it is the correct size, but when truncation mutants of ECSIT were made the band does not change size. SAMPLE Recombinant protein PRIMARY ANTIBODY Abcam ab21288, Diluted 1:1000 in TBST containing 2% BSA 0.01% NaN3. Incubated overnight at 4 degrees C. Followed by 3 quick washes, 3 15-min washes and 3 quick washes. DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Negative: Recombinant GFP and untransfected cells. Positive: Recombinant tagged ECSIT (of various lengths due to ECSIT truncation mutatants) ANTIBODY STORAGE CONDITIONS 4 Degrees C, as it arrived SAMPLE PREPARATION SDS Loading Buffer, 3 min at 95 degrees C AMOUNT OF PROTEIN LOADED 1 well of 24-well plate expressing protein. This is what I generally use for western blotting and works well for other combinations of recombinant proteins and respective antibodies. ELECTROPHORESIS/GEL CONDITIONS SDS PAGE, 4-12% Novex Bis Tris TRANSFER AND BLOCKING CONDITIONS For transfer: Invitrogen NuPage Transfer For Blocking: TBS-Tween (0.15M NaCl, 0.1 M Tris HCl pH7.4, 0.1% Tween 20) with 3% milk SECONDARY ANTIBODY [a competitor] goat anti-rabbit HRP conjugated 1:5000 Incubated 2 hours at 25 degrees C. Followed by 3 quick washes, 3 15-min washes and 3 quick washes. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Different cell lysates from independent transfections. Used fresh primary and secondary antibody dilutions every time. ADDITIONAL NOTES Uncertain which of the two is the order number: S102487 or 4500103431

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Answer

Thank you again for your enquiry. I spoke to a colleague of mine who also agreed that you may be detecting the endogenous protein. Did you cleave the tags? The tags may be adding the additional kDa’s to your bands. Another possibility is that differences in the size could be negligible enough that they do not register on your gel. There just is not enough information on your truncation methods to determine where the problem lies. Where is your truncated sample cleaved? The immunogen is within residues 301-350and I hope this information can help you. Please do not hesitate to contact us if you need any more advice or information.

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