beta-Defensin 3 Antibody - BSA Free

Novus Biologicals | Catalog # NB200-117

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Rat

Cited:

Human, Rat

Predicted:

Canine (90%), Porcine (90%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all beta-Defensin 3 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal portion of Defensin beta 3 (between amino acids 10-60) [UniProt P81534]

Reactivity Notes

Human and rat reactivity reported in (PMID: 21369366).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

7.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for beta-Defensin 3 Antibody - BSA Free

Immunohistochemistry-Paraffin: beta-Defensin 3 Antibody - BSA Free [NB200-117]

Immunohistochemistry-Paraffin: beta-Defensin 3 Antibody - BSA Free [NB200-117]

beta-Defensin-3-Antibody-Immunohistochemistry-Paraffin-NB200-117-img0002.jpg
Immunohistochemistry-Paraffin: beta-Defensin 3 Antibody - BSA Free [NB200-117]

Immunohistochemistry-Paraffin: beta-Defensin 3 Antibody - BSA Free [NB200-117]

Immunohistochemistry-Paraffin: beta-Defensin 3 Antibody [NB200-117] - Analysis of beta-Defensin 3 in human skin.
beta-Defensin 3 Antibody - BSA Free

Immunohistochemistry: beta-Defensin 3 Antibody - BSA Free [NB200-117] -

Immunohistochemistry: beta-Defensin 3 Antibody - BSA Free [NB200-117] - Topical application of PALA enhances AMP production in human skin explants. (a) HBD2 protein levels are increased in tissue homogenates of infected human skin explants by PALA or Neosporin treatment. Human skin biopsies were infected with MRSA for 1 h & then treated with the indicated ointments for 24 hours in triplicate. HBD2 levels were measured in tissue homogenates by ELISA (n = 3–4 donors). Mean ± SD; significance determined by 1-way ANOVA & Bonferroni multiple comparison test; *p < 0.05. (b) Immunohistochemistry staining of HBD2 in MRSA infected human skin explants described in (a). Representative image of 3 donors. (c) HBD3 protein levels in tissue homogenates of infected human skin explants are unaffected by PALA or Neosporin treatment. HBD3 levels were measured in tissue homogenates described in (a) by ELISA (n = 4 donors). Mean ± SD; significance determined by 1-way ANOVA & Bonferroni multiple comparison test; p > 0.05. (d) Immunohistochemistry staining of HBD3 in MRSA infected human skin explants described in (a). Representative image of 4 donors. (e) LL-37 protein levels are increased in tissue homogenates of infected human skin explants by PALA treatment. LL-37 levels were measured in tissue homogenates described in (a) by ELISA (n = 4–5 donors). Mean ± SD; significance determined by 1-way ANOVA & Bonferroni multiple comparison test; *p < 0.05. (f) Immunohistochemistry staining of LL-37 in MRSA infected human skin explants described in (a). Representative image of 6 donors. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29880914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for beta-Defensin 3 Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

reported in scientific literature

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:500-1:1000

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: beta-Defensin 3

A novel, nonhemolytic antimicrobial peptide (human beta-defensin-3, hBD-3) was isolated from human lesional psoriatic scales and clones from keratinocytes. hBD-3 demonstrated a salt-insensitive broad spectrum of potent antimicrobial activity against many potentially pathogenic microbes including multiresistant S. aureus and vancomyosin-resistant Entoerococcus faecium. Keratinocytes and airway epithelial cells are cellular sources of hBD-3. Tumor necrosis factor alpha and contact with bacteria were found to induce hBD-3 mRNA expression. hBD-3, therefore, might be important in the innate epithelial defense of infections by various microorganisms seen in skin and lung, such as cystic fibrosis.

Alternate Names

BD14, BD3, betaDefensin 3, DEFB103A, DEFB103B, DEFB3, HBD-3

Entrez Gene IDs

55894 (Human)

Gene Symbol

DEFB103B

UniProt

P81534

Additional beta-Defensin 3 Products

Product Documents for beta-Defensin 3 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for beta-Defensin 3 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for beta-Defensin 3 Antibody - BSA Free

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Protocols

View specific protocols for beta-Defensin 3 Antibody - BSA Free (NB200-117):


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


Western Blot Protocol
1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-Defensin beta 3 primary antibody (NB200-117) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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FAQs for beta-Defensin 3 Antibody - BSA Free

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  • Q: Are there by chance any WB images available for NB200-117 that haven’t been posted yet? If not, do you happen to know what band size(s) were actually detected? We have a customer who is concerned because he has encountered several products that are detecting around 15-17 kDa as opposed to the expected mw of ~8 kDa.

    A:

    We unfortunately do not have any images available for this product in western blot. I am sorry for any inconvenience. Since western blot is listed on our datasheet, we will of course fully guarantee the application. Typically a band is seen around 10kDa. Depending on the molecular weight marker, I would expect that there might be some difference seen at such a low molecular weight. 15-17kDa does seem a bit too high. I noticed that the protein does have several disulfide binding points. I would make sure your customer is properly boiling their samples (we recommend 10 minutes at 95C) and that they are adding at least 5% reducing agent to their sample buffer. We use BME typically, but DTT is also good.

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