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Product Info Summary
| SKU: | EK1514 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine®
View all alpha-Defensin 1 ELISA kits
SKU/Catalog Number
EK1514
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine® (96 Tests). Quantitate Human DEFA1 in cell culture supernatants, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1514)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: D39-C94
Sensitivity
<10 pg/ml
Assay Range
312 pg/ml - 20,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1514 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1514 is reactive to DEFA1 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA).
Application Guarantee
EK1514 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of alpha-Defensin 1
Defensin is a human protein that is encoded by the DEFA1 gene. Defensins are a family of antimicrobial and cytotoxic peptides thought to be involved in host defense. They are abundant in the granules of neutrophils and also found in the epithelia of mucosal surfaces such as those of the intestine, respiratory tract, urinary tract, and vagina. The protein encoded by this gene, defensin, alpha 1, is found in the microbicidal granules of neutrophils and likely plays a role in phagocyte-mediated host defense. Members of the defensin family are highly similar in protein sequence and distinguished by a conserved cysteine motif. Several alpha defensin genes are clustered on chromosome 8. This gene differs from defensin, alpha 3 by only one amino acid. This gene and the gene encoding defensin, alpha 3 are both subject to copy number variation.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1514-CAP | Anti-Human DEFA1 Pre-coated 96-well strip microplate | 1 |
| EK1514-ST | Human DEFA1 Standard | 2 vials, 10 ng/tube |
| EK1514-DA | Human DEFA1 Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 312 | 625 | 1250 | 2500 | 5000 | 10000 | 20000 |
| O.D. | 0.091 | 0.139 | 0.172 | 0.203 | 0.353 | 0.633 | 1.277 | 1.963 |
Data Example Images
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Human alpha-defensin/DEFA1 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect alpha-Defensin 1, For serum, dilution ratio is 1:1000 and concentration is around 2.5 ug/ml.
For plasma, dilution ratio is 1:10000 and concentration is around 30 ug/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 462 | 1682 | 9496 | 635 | 1897 | 9984 |
| Standard deviation | 21.71 | 89.15 | 531.78 | 42.55 | 96.75 | 479.23 |
| CV (%) | 4.7% | 5.3% | 5.6% | 6.7% | 5.1% | 4.8% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of alpha-Defensin 1 in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 462 | 429 | 405 | 424 | 430 | 20.53 | 4.7% |
| Sample 2 | 1682 | 1777 | 1969 | 1748 | 1794 | 106.74 | 5.9% |
| Sample 3 | 9496 | 8586 | 8746 | 8590 | 8854 | 375.94 | 4.2% |
Protein Target Info & Infographic
Gene/Protein Information For DEFA1 (Source: Uniprot.Org, NCBI)
Gene Name
DEFA1
Full Name
Neutrophil defensin 1
Weight
10.201kDa
Superfamily
alpha-defensin family
Alternative Names
Neutrophil defensin 1; Defensin, alpha 1; HNP-1; HP-1; HP1; HP 1-56; Neutrophil defensin 2; HNP-2; HP-2; HP2; DEFA1; DEF1; DEFA2; MRS; DEFA1B DEFA1 DEF1, DEFA2, HNP-1, HP-1, HP1, MRS defensin alpha 1 neutrophil defensin 1|defensin, alpha 1, myeloid-related sequence|defensin, alpha 2|myeloid-related sequence
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on DEFA1, check out the DEFA1 Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for DEFA1: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine® (EK1514)
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Customer Q&As
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11 Customer Q&As for Human alpha-defensin 1/DEFA1 ELISA Kit PicoKine®
Question
Can EK1514 be used on synovial fluid?
Verified customer
Asked: 2020-10-26
Answer
Our lab technicians haven't tested the Human Alpha-Defensin/DEFA1 ELISA Kit PicoKine™ (EK1514) on synovial fluid. We suggest running a pilot test. In theory, the kit is able to detect the target protein if the expression level of DEFA1 in the sample falls in the assay range of EK1514.
Boster Scientific Support
Answered: 2020-10-27
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
W. Adams
Verified customer
Asked: 2020-08-10
Answer
A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are included in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2020-08-10
Question
Does EK1514 cross reacts with other defensin proteins. Also, what other proteins have been tested for cross-reactivity?
Verified Customer
Verified customer
Asked: 2020-04-28
Answer
The following recombinant proteins were tested for cross-reactivity: <br> <br> Recombinant human: α-Defensin 5, β-Defensin 3, C1qB, MCP-2 and Serpin F2. No significant cross reactivity was observed.
Boster Scientific Support
Answered: 2020-04-28
Question
Q: What is the optimal O.D. value for alpha-Defensin 1 ELISA kit? I used your alpha-Defensin 1 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain alpha-Defensin 1 even though the O.D. values are not very high?
A. Wilson
Verified customer
Asked: 2019-06-21
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. what you should focus on is whether your sample O.D. values are statistically significantly higher than your blank values. regarding your assay, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this alpha-Defensin 1 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of alpha-Defensin 1 that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-06-21
Question
Can the reconstituted stock standard solutions of EK1514 be stored for further used? How can it be stored and for how long?
Verified customer
Asked: 2019-04-16
Answer
For the Human Alpha-Defensin/DEFA1 ELISA Kit PicoKine™ (EK1514), our lab technicians suggests using a fresh standard for each assay. The reconstituted stock standard solutions can be stored for further use, although not for a long time. The reconstituted standard can be stored at -20°C for 48 hours and 12 hours at 4°C.
Boster Scientific Support
Answered: 2019-04-16
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring alpha-Defensin 1 using the EK1514 Human alpha-Defensin 1 Picokine® ELISA Kit?
C. Brown
Verified customer
Asked: 2019-01-15
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples to avoid protein degradation.
Boster Scientific Support
Answered: 2019-01-15
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2018-08-22
Answer
A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.
Boster Scientific Support
Answered: 2018-08-22
Question
Q: which procedure should I follow in order to thaw whole blood sample for alpha-Defensin 1 ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-03-27
Answer
A: we do not recommend freezing and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test alpha-Defensin 1 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.
Boster Scientific Support
Answered: 2018-03-27
Question
Will the antibodies used in the kit react to HNP2 and HNP3?
Verified Customer
Verified customer
Asked: 2016-11-07
Answer
The antibodies used in the kit are predicted to react with HNP2 and HNP3.
Boster Scientific Support
Answered: 2016-11-07
Question
Q: is it okay to use citrate plasma as samples in Human alpha-Defensin 1 Picokine® ELISA Kit (Catalog # EK1514)?
Verified Customer
Verified customer
Asked: 2016-08-27
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of alpha-Defensin 1 causing disruption of its protein structure. alpha-Defensin 1 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the alpha-Defensin 1 ELISA, treating samples with different anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2016-08-27
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2014-03-09
Answer
A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2014-03-09
