Overview

  • Product name
    Anti-BD-3 antibody [L3-18b-E1]
    See all BD-3 primary antibodies
  • Description
    Mouse monoclonal [L3-18b-E1] to BD-3
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, RIA, IHC-Frmore details
  • Species reactivity
    Reacts with: Human, Chimpanzee
  • Immunogen

    Synthetic peptide corresponding to Human BD-3 aa 6-22.
    Sequence:

    LQKYYCRVRGGRCAVLS


    (Peptide available as ab49530)

  • General notes

    Previously labelled as beta Defensin 3. 

Properties

Applications

Our Abpromise guarantee covers the use of ab14422 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
ELISA Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 21056450

Target

  • Function
    Exhibits antimicrobial activity against Gram-positive bacteria S.aureus and S.pyogenes, Gram-negative bacteria P.aeruginosa and E.coli and the yeast C.albicans. Kills multiresistant S.aureus and vancomycin-resistent E.faecium. No significant hemolytic activity was observed.
  • Tissue specificity
    Highly expressed in skin and tonsils, and to a lesser extent in trachea, uterus, kidney, thymus, adenoid, pharynx and tongue. Low expression in salivary gland, bone marrow, colon, stomach, polyp and larynx. No expression in small intestine.
  • Sequence similarities
    Belongs to the beta-defensin family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • BD 3 antibody
    • BD-3 antibody
    • BD3 antibody
    • beta 103 antibody
    • Beta defensin 103A antibody
    • Beta defensin 103A precursor antibody
    • Beta defensin 3 antibody
    • Beta-defensin 103 antibody
    • Beta-defensin 3 antibody
    • D103A_HUMAN antibody
    • DEF B3 antibody
    • DEFB 103 antibody
    • DEFB 103A antibody
    • DEFB 3 antibody
    • DEFB-3 antibody
    • DEFB103 antibody
    • DEFB103A antibody
    • DEFB103B antibody
    • DEFB3 antibody
    • Defensin antibody
    • Defensin beta 103A antibody
    • Defensin beta 3 antibody
    • Defensin like protein antibody
    • Defensin-like protein antibody
    • HBD 3 antibody
    • hBD-3 antibody
    • HBD3 antibody
    • HBP 3 antibody
    • HBP3 antibody
    • mBD3 antibody
    see all

References

This product has been referenced in:
  • Yost S  et al. Potassium is a key signal in host-microbiome dysbiosis in periodontitis. PLoS Pathog 13:e1006457 (2017). IHC . Read more (PubMed: 28632755) »
  • Tugizov SM  et al. HIV is inactivated after transepithelial migration via adult oral epithelial cells but not fetal epithelial cells. Virology 409:211-22 (2011). IHC-Fr ; Human . Read more (PubMed: 21056450) »
See all 2 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Following up my previous message from Wednesday, I have received the following protocol from the laboratory that valdiated this antibody for ELISA: We tested the antibody in ELISA with the following conditions: Coating of plate with 100 µl of (2 µg/ml) human beta-Defensin 3 (aa 6-22) in bicarbonate buffer, Blocking with BSA Detection with ab14422 in serial dilutions ranging from 1:250 to 1:1000 gave a good signal Substrate: TMB Is this similar to what you tried? We are confident that what you received is mouse anti-BD3, but the third ELISA experiment results that you described earlier suggests that what you received is defective, if your protocol approximated these conditions, and if your secondary antibody was anti-mouse IgG.

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Answer

Thank you for the datasheets of the secondary antibodies. I am surprised that Thermo recommended 31462. The datasheet for this secondary includes this note: The product has been tested by ELISA and/or solid-phase adsorbed to ensure minimal cross-reaction with human serum proteins. However, this antibody may cross-react with immunoglobulins from other species. This is in the "Antibody Specificity" field of the datasheet. You may want to ask them about that, because it suggests that reactivity with mouse IgG was not tested. I will find out from the laboratory that produced the antibody how it was validated for ELISA and will forward this information to you. If you can confirm that your protocol is similar, with regards to the concentration of peptide and antibody, and the protocol, I will be happy to send a replacement or refund. I assure you I want to resolve this quickly, hopefully before the end of the week.

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Answer

Thank you for the clarification. I have just  a few more questions. 1. In the first experiment, did the anti-mouse secondary react with ab14422? 2. What is the source and catalogue number for the anti-rabbit secondary that reacted with ab14422? 3. For your second experiment, did you test in parallel any other mouse IgG, in addition to ab14422? What is the source and catalogue numbers of the two anti-rabbit antibodies? I understand you have not seen this before with your anti-rabbit secondaries, but polyclonal secondaries can vary lot to lot with regards to species reactivity. 4. In the third experiment, was the peptide that the plate was coated with the same as the peptide used as the immunogen for ab14422, LQKYYCRVRGGRCAVLS, corresponding to amino acids 6-22 of Human beta 3 Defensin? What is the control antibody in this experiment? What is the secondary? We will not need the data, as you have described the issue well. At this point, I still suspect the specificity of your anti-rabbit secondaries. On the otther hand, if the peptide coated onto the plate contains the ab14422 immunogen sequence, and the anit-mouse IgG secondary fails to detect ab14422, and is not isotype subclass-specific (for example, anti- IgG2a), then I do suspect ab14422. I look forward to your reply.  

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Answer

Thank you for the clarification. We are confident that ab14422 is a mouse monoclonal antibody. Have you used the secondary with other mouse antibodies, isotype IgG1? That is the isotype (IgG1) of this clone (L3-18b-E1). I recommend contacting the company that produces the secondary. They may have mouse IgG cross-reactivity data for the lot you have. If you need an anti-rabbit secondary that will not react with mouse IgG1, I will be happy to recommend one from our catalogue. We have several anti-rabbit antibodies that are pre-adsorbed to remove reactivity with mouse IgG and other species, with several different conjugations, for example ab96899: Click here (or use the following: https://www.abcam.com/index.html?datasheet=96899).

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Answer

Thank you for contacting us. The BD-3 antibody is a mouse monoclonal antibody. Is the anti-rabbit secondary also an Abcam antibody? If it has not been pre-adsorbed to remove reactivity with mouse IgG, it is possible it cross-reacts with mouse IgG, even though it is raised against rabbit IgG, since immunoglobulins are conserved to varying extent across many species.

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Answer

Thank you for getting back to me with those details, it was very interesting. Unfortunately given that the source of the antisera have not been able to provide me with further details as to the synthesis of the immunising peptides it is difficult for me to comment on whether they have been raised against a correctly folded immunogen. If this was indeed the case then one might speculate that these antibodies only recognise the synthetic peptide and not the native form of the protein. However, checking back to past orders and this antibody is relatively popular and we have not received any complaints as such. I can but speculate that the absence of western or dot blot as applications on the datasheets is a reflection that these antibodies do not recognise their epitopes when immobilised on a membrane.

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Answer

Thank you for your enquiry. Further to correspondence with the source of this antiserum I have been informed that the antiserum was raised against tricycle human ß-Defensin 2 antigen. Unfortunately they could not provide details of whether the antisera had been tested against the native protein or your protein standards. I have therefore not been able to find evidence that these sera have been tested against the native form of this protein. I would definitely be interested to know how these sera performed against the denatured forms of your standards.

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Answer

Thank you for contacting me. Unfortunately I have not been able to obtain a satisfactory answer from the source of this antiserum as yet. I will be in touch as soon as I know more details about the synthesis of the immunising peptides. I appreciate your patience.

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