Overview

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Constituent: Whole serum
  • Purity
    Whole antiserum
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab14421 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.
WB Use at an assay dependent concentration. Predicted molecular weight: 8 kDa.
IHC-Fr Use at an assay dependent concentration. PubMed: 21056450

Target

  • Function
    Has bactericidal activity.
  • Tissue specificity
    Plasma.
  • Sequence similarities
    Belongs to the beta-defensin family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • BD 1 antibody
    • BD-1 antibody
    • BD1 antibody
    • beta 1 antibody
    • Beta defensin 1 antibody
    • Beta-defensin 1 antibody
    • DEFB 1 antibody
    • DEFB1 antibody
    • DEFB1_HUMAN antibody
    • DEFB101 antibody
    • Defensin antibody
    • Defensin beta 1 antibody
    • Defensin beta 1 preproprotein antibody
    • HBD 1 antibody
    • hBD-1 antibody
    • HBD1 antibody
    • MGC51822 antibody
    see all

References

This product has been referenced in:
  • Pierson T  et al. Cigarette smoke extract induces differential expression levels of beta-defensin peptides in human alveolar epithelial cells. Tob Induc Dis 11:10 (2013). IF . Read more (PubMed: 23627872) »
  • 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

1-4 of 4 Abreviews or Q&A

Answer

Thank you for your enquiry. Human beta-1 defensin is expressed in kidney. Therefore, I would suggest you can use a human kidney whole cell lysate such as our peptide product ab30203. I have placed a link here to a reference I hope you may find useful from BMC Infectious Diseases; Expression of human beta-defensins 1 and 2 in kidneys with chronic bacterial infection (Jan Lehmann1 et al) http://www.biomedcentral.com/content/pdf/1471-2334-2-20.pdf Alternatively, you could use a purified beta-1 defensin peptide/protein. I hope this is helpful. Please do not hesitate to contact us again if you require more assistance.

Read More

Question

My colleague, wrote you about problem he is having with an antibody we bought from you, and we got a reply from you (please see below for the original message and your reply). In sum, we bought your anti-beta-1 definsin antibody (AB14421) based on your advertisement recommending this antibody for ELSIA detection of beta-1 defensin in human samples. In our tests using a commercial purified beta-1 defensin in our ELISA assay, we never were able to detect anything with your antibody. In previous email communications, we raised this issue and you provided a complete protocol for the ELISA assay. We followed the protocol and still were not able to detect anything. We followed up by asking for your clarification on the source of the antigen in your assays. In your reply below you now tell us that your ELISA was tested positive on a synthetic peptide generated based on a partial sequence of human beta-1 defensin. Moreover the synthetic peptide contains several amino acid changes from human beta-1 defensin. It is wonderful your antibody can detect this synthetic peptide. However, for you to advertise the antibody for beta-1 defensin ELISA, do you have any data supporting this claim: do you have anything you show me to confirm the antibody can actually detect human beta-1 defensin (native, synthetic or recombinant form of the entire protein)? Obviously, the only reason we (and others) ended up buying this antibody is for use in beta-1 defensin detection, not detection of a synthetic peptide. If you have data that your antibody can detect intact human beta-1 defensin, can you give me detail how this is done (e.g. source of defensin, how much defensin, condition used, and result of dose-response curve). If none of this information is available, please advise what we need to do with the item we bought from you based on what you advertised. Thank you in advance for your prompt attention to this matter.

Read More
Answer

Thank you very much for your enquiry. I have contacted the originator of this antibody, and they tested the antibody in ELISA using human beta Defensin 1 (aa1-36) (DHYNCVSSGGQCLYSACPIFTKIQGTCYRGKAKCCK). In this test the antibody had a titer of 1 to 3000. The peptide against which the antibody was made, was a synthetic peptide: YRGKAKC(ACM)G, corresponding to amino acids 28-34 of Human beta 1 Defensin. As your colleague has reported, no signal was obtained using ab14421 in ELISA. We would very much like to investigate this problem, but I do need require some information regarding your protocol. If you could kindly answer the questions below, we will do our best to determine what is going on. Thank you, and I look forward to hearing from you. Also - what was the batch number that you received (it is located on the vial)? 1. Please describe the problem (high background, no signal, non-specific colour development, poor standard curve etc). 2. What type of ELISA are you performing? (Direct ELISA, indirect ELISA, sandwich ELISA etc.) 3. On what material are you testing the antibody in ELISA? •Species? •Cell type? •etc? 3. How did you coat the plates? 4. How did you block the unspecific binding sites? For how long? How did you wash the wells? 5. Primary antibody •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 6. Secondary antibody •What secondary antibody are you using? •Specification (in which species was it raised against)? •At what dilution(s) have you tested this antibody? •Incubation, wash steps? •Do you know whether the problems you are experiencing come from the secondary? 7. Which detection system did you use? Substrate? 8. Did you apply positive and negative controls along with the samples? Please specify. What did you use for standards? 9. Optimization attempts •How many times have you tried the ELISA? •Do you obtain the same results every time? •What steps have you altered?

Read More

Answer

Thank you for your email. We source ab14421 from outside Abcam and the originator used a synthetic peptide with the sequence: Human b-Defensin 1 (aa 28-34) (YRGKAKCACMG) (underlined amino acids differ from the original sequence, they were added or substituted for technical reasons). They used the antibody at a dilution of 1:1000. If you need further assistance, please let me know.

Read More

Question
Answer

Below is the ELISA protocol which the originator of ab14421 followed. If you have any additional questions, please contact us again. PROTOCOL DIRECT ELISA: REAGENTS: Coating buffer: 0,05 M Na2CO3 adjust pH to 9,6 with solid NaHCO3 Blocking buffer: 2% BSA in PBS Washbuffer: 1,37 Nacl, 26,8 mM KCl, 14,7mM KH2PO4, 81 mM Na2HPO4, 1 % TRITON-X-100, 0,02% THIMEROSAL First antibody: dilute in 2% BSA in PBS Second antibody: dilute in wash buffer Substrate : TMB-solution ( BIOFX, USA) Stop solution: 0,4 M H2SO4 Microtiterplate NUNC Medisorb PROCEDURE: · pipette 100 µl of peptide (500 ng/ml in coating buffer) into the wells of a microtitre plate · incubate over night at 4 °C · decant the contents of the plate and add 250 µl blocking buffer · incubate 1 hour at room temperature, shaking on a horizontal mixer · decant the content of the plate and wash the cavities 5 x with 250 µl of wash buffer · pipette 100 µl of antiserum (respective dilution in 2% BSA / PBS) · incubate at least 3 hrs at room temperature (or over night at 4 °C), shaking on a horizontal mixer · decant the content of the plate and wash the cavities 5 x with 250 µl of wash buffer · pipette 100 µl of POX-labelled second antibody (e.g. if first antibody is rabbit: donkey antirabbit- POX from Dako can be used at 1: 10000 in wash buffer) · incubate 1 hr at room temperature, shaking on a horizontal mixer · decant the content of the plate and wash the cavities 5 x with 250 µl of wash buffer · pipette 100 µl TMB-solution · incubate for 15 - 30 minutes at room temperature, shaking slightly, until sufficient color differentiation has been observed · add 50 µl of stop solution and mix shortly · measure the extinction of the samples at 450 nm (reference wave length 620 nm)

Read More

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

Sign up