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Product Info Summary
| SKU: | EK1469 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum, plasma (heparin, EDTA) , saliva, urine and human milk. |
Product info
Product Name
Human HE4 ELISA Kit PicoKine®
SKU/Catalog Number
EK1469
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human HE4 ELISA Kit PicoKine® (96 Tests). Quantitate Human WFDC2 in cell culture supernatants, serum, plasma (heparin, EDTA) , saliva, urine and human milk. Sensitivity: 15pg/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 HE4 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1469)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: E.coli; Immunogen sequence: E31-F124
Sensitivity
<15 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1469 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1469 is reactive to WFDC2 in Human samples
Validated Sample Types
cell culture supernatants, serum, plasma (heparin, EDTA) , saliva, urine and human milk.
Application Guarantee
EK1469 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of HE4/WFDC2
WAP four-disulfide core domain protein 2, also known as Human Epididymis Protein 4 (HE4), is a protein that in humans is encoded by the WFDC2 gene. This gene encodes a protein that is a member of the WFDC domain family. The WFDC domain, or WAP Signature motif, contains eight cysteines forming four disulfide bonds at the core of the protein, and functions as a protease inhibitor in many family members. This gene is expressed in pulmonary epithelial cells, and was also found to be expressed in some ovarian cancers. The encoded protein is a small secretory protein, which may be involved in sperm maturation.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1469-CAP | Anti-Human WFDC2 Pre-coated 96-well strip microplate | 1 |
| EK1469-ST | Human WFDC2 Standard | 2 vials, 10 ng/tube |
| EK1469-DA | Human WFDC2 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 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.044 | 0.231 | 0.379 | 0.594 | 0.929 | 1.304 | 1.650 | 1.894 |
Data Example Images
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Human HE4 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect HE4/WFDC2, Dilution ratio of 1:50, concentration in serum and plasma is 31.4-41.1 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human HE4 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) | 246 | 1139 | 4402 | 255 | 1184 | 4098 |
| Standard deviation | 14.02 | 77.45 | 176.08 | 18.62 | 101.82 | 176.21 |
| CV (%) | 5.7% | 6.8% | 4% | 6.4% | 8.6% | 4.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of HE4/WFDC2 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 | 246 | 276 | 254 | 281 | 264 | 14.63 | 5.5% |
| Sample 2 | 1139 | 1088 | 1273 | 1192 | 1173 | 68.45 | 5.8% |
| Sample 3 | 4402 | 3964 | 4189 | 4282 | 4209 | 160.47 | 3.8% |
Protein Target Info & Infographic
Gene/Protein Information For WFDC2 (Source: Uniprot.Org, NCBI)
Gene Name
WFDC2
Full Name
WAP four-disulfide core domain protein 2
Weight
12993 MW
Alternative Names
WAP four-disulfide core domain protein 2;Epididymal secretory protein E4;Major epididymis-specific protein E4;Putative protease inhibitor WAP5;WFDC2;HE4, WAP5; WFDC2 EDDM4, HE4, WAP5, dJ461P17.6 WAP four-disulfide core domain 2 WAP four-disulfide core domain protein 2|WAP domain containing protein HE4-V4|epididymal protein 4|epididymal secretory protein E4|epididymis secretory sperm binding protein|epididymis-specific, whey-acidic protein type, four-disulfide core|human epididymis protein 4|major epididymis-specific protein E4|putative protease inhibitor WAP5
*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 WFDC2, check out the WFDC2 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 WFDC2: 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 HE4 ELISA Kit PicoKine® (EK1469)
Hello CJ!
EK1469 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Value of HE4 Combined with Cancer Antigen 125 in the Diagnosis of Endometrial Cancer
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Customer Reviews
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Customer Q&As
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12 Customer Q&As for Human HE4 ELISA Kit PicoKine®
Question
Does EK1469 contain native human biological materials?
Verified customer
Asked: 2021-11-10
Answer
The Human HE4 ELISA Kit PicoKine® (EK1469) doesn't contain native human biological materials. doesn't contain native human biological materials.
Boster Scientific Support
Answered: 2021-11-11
Question
Q: which procedure should I follow in order to thaw whole blood sample for HE4 ELISA after freezing?
Verified Customer
Verified customer
Asked: 2020-08-12
Answer
A: do not freeze 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 HE4 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: 2020-08-12
Question
Q: we need your recommendation regarding the dilution ratio of serum samples for detection of HE4 in Human human milk? I am trying to measure a few analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.
B. Mehta
Verified customer
Asked: 2020-06-22
Answer
A: unable to understand the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the HE4 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.
Boster Scientific Support
Answered: 2020-06-22
Question
What is the dilution ratio for urine samples with your in-house testing of the EK1469 kit?
Verified customer
Asked: 2020-01-02
Answer
The dilution ratio for urine samples using the Human HE4 ELISA Kit PicoKine™ EK1469 is 1:200. The sample values might be varied with urine samples from different people.
Boster Scientific Support
Answered: 2020-01-13
Question
Q: What is the optimal O.D. value for HE4 ELISA kit? I performed your HE4 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 HE4 even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-12-02
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. considering 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. typically, 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 changeing 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 HE4 ELISA kit to have sensitivity of 15pg/ml, that means the minimum amount of HE4 that can be declared/interpreted as positive by the above standard is 15pg/ml.
Boster Scientific Support
Answered: 2019-12-02
Question
Q: if the enzyme conjugated HE4 antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the HE4 antigen?
L. Mangal
Verified customer
Asked: 2019-06-23
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2019-06-23
Question
Q: how many samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2018-11-24
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 might differ by kit so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2018-11-24
Question
Q: Can HE4 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2018-10-10
Answer
A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is required. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please see the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.
Boster Scientific Support
Answered: 2018-10-10
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2018-08-22
Answer
A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions
Boster Scientific Support
Answered: 2018-08-22
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
L. Zhou
Verified customer
Asked: 2018-04-27
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: 2018-04-27
Question
What is the conversion factor for He4 to transfer units from pg/ml to pmol/l?
Verified Customer
Verified customer
Asked: 2018-04-05
Answer
The conversion factor for He4 to transfer units from pg/ml to pmol/l is 0.077, which means 1pg/ml euquals to 0.077pmol/l.
Boster Scientific Support
Answered: 2018-04-05
Question
Q: is it okay to use citrate plasma as samples in Human HE4 Picokine® ELISA Kit (Catalog # EK1469)?
G. Liu
Verified customer
Asked: 2015-08-13
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of HE4 causing degradation of its protein structure. HE4 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the HE4 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: 2015-08-13
