Overview

  • Product name
  • Description
    Rabbit polyclonal to BD-3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human BD-3.

  • General notes

    Previously labelled as beta Defensin 3. 

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab19270 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
WB 1/500 - 1/1000. Detects a band of approximately 5 kDa (predicted molecular weight: 5 kDa).
ICC/IF Use at an assay dependent concentration. PubMed: 23627872

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

Images

  • ab19270 staining BD-3 in the A549 cell line from Human Lung by ICC/IF (Immunocytochemistry/immunofluorescence) treated with cigarette smoke extract. Cells were fixed , permeabilized and blocked. Samples were incubated with primary antibody (1/500) for 1 hour. An  Alexa Fluor®488-conjugated  was used as the secondary antibody and DAPI used to stain the nuclei.

References

This product has been referenced in:
  • Miani M  et al. Gut Microbiota-Stimulated Innate Lymphoid Cells Support ß-Defensin 14 Expression in Pancreatic Endocrine Cells, Preventing Autoimmune Diabetes. Cell Metab N/A:N/A (2018). Read more (PubMed: 30017352) »
  • Zhu M  et al. Expression and antimicrobial character of cells transfected with human ß-defensin-3 against periodontitis-associated microbiota in vitro. Mol Med Rep 16:2455-2460 (2017). Read more (PubMed: 28677765) »
See all 4 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Sure, I am happy to send along the replacements.

If you could please remind me of the purchase order number or Abcam order reference number associated with the original purchase of these antibodies, I'll arrange replacements immediately.

Thanks!

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Equus Tissue sections (respiratory tract)
Permeabilization
Yes - 0.1% Triton X 100
Specification
respiratory tract
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde

Jolien Van Cleemput

Verified customer

Submitted Dec 07 2015

Answer

Does your customer want to use the same catalog numbers or would she be interested in choosing one or more different antibodies (of the same price) to the beta defensin proteins?

Read More

Answer

Thanks for passing along the peptide and protein concentration information.

These concentrations seem reasonable.

Is your customer interested in replacements for the antibodies?

Read More

Answer

Thank you for sending along the dot blots and additional information. We really appreciate the extra troubleshootingeffort.

Here are my thoughts:

1. In general it looks like the antibodies are able to detect Defensin in the chorioamniotic membrane samples.

2. It does not appear that the antibodies detected recombinant Defensin protein. You note the mass of recombinant protein spotted on the membranes, from 400ng - 0.78ng. Was the concentration of the Defensin proteinconfirmed by Bradford assay or OD280n measurements following reconstitution or are these number based on the expected concentrations after reconstitution?

If the concentrations are only theoretical, could you please check the concentrations to ensurethey are accurate?

Thanks again for your troubleshooting efforts. I'm sure we'll be able to resolve this situation soon.

Read More

Question

For the technical support,   Please, customer is having problems using human beta defensin antibodies and their respective recombinant proteins in WB applications.   Find bellow the technical questionnaire filled out by the customer.   As the beta defensins are small proteins (8 kDa), it seems this might be the origin of the problem. Maybe, the customer is losing these small bands during the running or even during the transfer. Could you please verify her protocol and give suggestions on how to solve this problem?   Thank you in advance.   Kind Regards, Abcam WB Questionnaire   1) Abcam product code ab14425 + ab50048 ab66072 + ab9872 ab19270 + ab50059 ab14419 + ab69499                            2) Lot number ab14425: GR 17872-2 + ab50048: GR 45073-1 ab66072: GR 40785-1 + ab9872: GR 11654-6 ab19270: GR 730-1 + ab50059: GR 1609-2 ab14419: GR 21054-1 + ab69499: GR 3598-2     3) Antibody storage conditions (temperature/reconstitution etc) Antibodies: stored at -20 ºC. Recombinant proteins: after adding 100 ul PBS + 15% glycerol we aliquoted each one and stored at -80°C.   4) Description of the problem (high background, wrong band size, more bands, no band etc.) No bands using the beta defensin antibodies with their respective recombinant protein and no bands with lysate of chorioamniotic membrane.   5) Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Human recombinant proteins and lysate of chorioamniotic membrane tissue from human.   6) Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Lysis buffer: - 50mM Tris HCl pH 7,4 - 0,2mM NaCl - 0,1% Triton X-100 - 10mM CaCl2 - 10ul/mL Protease inhibitor from GE Healthcare   Heating sample for 5 min at 100ºC.     7) Amount of protein loaded 30ug   8) Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) 15% gel Running at 100 V for 1h30min Sample buffer (4x): - 250mM Tris-HCl 0,5M pH:6,8 - 8% SDS - 20% B-mercaptoethanol - 0,012g bromophenol blue -100 mL milli-Q water - 30% glycerol     9) Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) Transfer: 1h30min at 120V Transfer buffer: 9,7g Tris, 57,6g glycine, 3,2L distilled water and 0,8L methanol Transfer membranes tested: 0,45µm Hybond membrane from Amersham and 0,22µm membrane from Millipore.   Blocking conditions tested: 5% non-fat Milk diluted in TBS-T for 1h at room temperature under agitation. 5% non-fat Milk diluted in TBS-T for 2h at room temperature under agitation. 10% non-fat Milk diluted in TBS-T for 1h at room temperature under agitation. 10% non-fat Milk diluted in TBS-T for 2h at room temperature under agitation.   10) Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) 1. Human Beta Defensin-1:  ab14425 (antibody) + ab50048 (rec protein) Tested dilutions: 1:500 and 1:250 for the antibody and 200 ng for the recombinant protein   2. Human Beta Defensin-2: ab66072 (antibody) + ab9872 (rec protein) Tested dilutions: 1:1000 and 1:500 for the antibody and 200 ng for the recombinant protein   3.  Human Beta Defensin-3: ab19270 (antibody) + ab50059 (rec protein) Tested dilutions: 1:1000 and 1:500 for the antibody and 200 ng for the recombinant protein   3.  Human Beta Defensin-4: ab14419 (antibody) + ab69499 (rec protein) Tested dilutions: 1:500 and 1:250 for the antibody and 200 ng for the recombinant protein   All antibodies were diluted in TBST + 5% milk and incubated overnight at 4°C under agitation. We also tried to dilute them in TBST + 5% BSA.   Wash step: 3x 5 min in TBST   11) Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Goat anti-mouse IgG HRP for ab14425, ab66072 and ab14419. Dilution: 1:5000 diluted in TBST +5% milk Incubation: 1 h   Goat anti-rabbit IgG HRP F(ab’) fragment for ab19270. Dilution: 1:5000 diluted in TBST +5% milk Incubation: 1 h   Wash step: 3x 5 min in TBST   12) Detection method (ECL, ECLPlus etc.) ECL Plus - Amersham   13) How many times have you tried the Western? 20 times   14) Do you obtain the same results every time? e.g. are the background bands always in the same place? The results we have are always the same for the 4 proteins. The respective bands of recombinant protein and sample do not appear. The running and transfer conditions are ok, because we have positive results with beta actin antibody in the same membranes. Also, the secondary antibody is working fine with beta actin.   15) What steps have you altered to try and optimize the use of this antibody? Primary antibody dilution, transfer membranes, blocking conditions...      

Read More
Answer

Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is are having problems with 8 of our products.  Special thanks for collecting the troubled customer's data/results in such an organized way. It may well be that the problem is either shipment/storage or protocol-related. Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.   1) Arrival dates: Could you please confirm the exact dates when the customer received these items: - ab14425, ab50048, ab66072, ab9872, ab19270, ab50059, ab14419, ab69499 2) Orders and shipment: - Could you provide me the Abcam Order Numbers for these products? - Were these items shipped in the same package or not? 3) Marker bands: - What MW markers (i.e. range of the proteins for MW) were used for WB? I am particularly interested in the markers at the lower weight range. 4) Image: - Could you please attach an image representing the samples and the loading control on the same gels? 5) Secondary antibodies: - Has the customer used the respective secondary antibodies successfully with other primary antibodies? Could you please check if the problem does not come from the 2ndaries?   Thank you for your understanding and co-operation in this matter. I look forward to hearing from you and hope to solve this problem as soon as possible.

Read More

Answer

Thank you for your enquiry. The immunogen was recombinant human beta-defensin-3. This same material was used in Western blots using an SDS-PAGE mini gel. I have updated the datasheet to reflect that the immunogen is recombinant. Please let us know if you have any further questions.

Read More

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

Sign up