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Product Info Summary
| SKU: | EK1536 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum, plasma(heparin, EDTA), cell lysates and urine. |
Product info
Product Name
Human Nectin-4/PVRL4 ELISA Kit PicoKine®
View all Nectin-4/PVRL4 ELISA kits
SKU/Catalog Number
EK1536
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Nectin-4/PVRL4 ELISA Kit PicoKine® (96 Tests). Quantitate Human NECTIN4 in cell culture supernatants, serum, plasma(heparin, EDTA), cell lysates and urine. 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 Nectin-4/PVRL4 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1536)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: G32-S349
Sensitivity
<10 pg/ml
Assay Range
31.2 pg/ml - 2,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1536 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1536 is reactive to NECTIN4 in Human samples
Validated Sample Types
cell culture supernatants, serum, plasma(heparin, EDTA), cell lysates and urine.
Application Guarantee
EK1536 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Nectin-4/PVRL4
Nectin-4, also known as PVRL4, is an adhesion receptor of the immunoglobulin superfamily and functions in cell-cell adhesion. This gene is mapped to chromosome 1q23.3. The encoded protein contains two immunoglobulin-like (Ig-like) C2-type domains and one Ig-like V-type domain. It is involved in cell adhesion through trans-homophilic and -heterophilic interactions. It is a single-pass type I membrane protein. The soluble form is produced by proteolytic cleavage at the cell surface by the metalloproteinase ADAM17/TACE. The secreted form is found in both breast tumor cell lines and breast tumor patients. Mutations in this gene are the cause of ectodermal dysplasia-syndactyly syndrome type 1, an autosomal recessive disorder. Alternatively spliced transcript variants have been found but the full-length nature of the variant has not been determined.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1536-CAP | Anti-Human NECTIN4 Pre-coated 96-well strip microplate | 1 |
| EK1536-ST | Human NECTIN4 Standard | 2 vials, 10 ng/tube |
| EK1536-DA | Human NECTIN4 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 | 31.25 | 62.5 | 125 | 250 | 500 | 1000 | 2000 | |
| O.D. | 0.026 | 0.141 | 0.257 | 0.469 | 0.82 | 1.336 | 1.832 | 2.182 |
Data Example Images
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Human Nectin-4 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Nectin-4/PVRL4, For serum, dilution ratio is 1:1 and concentration is 80-500 pg/ml.
For plasma, dilution ratio is 1:1 and concentration is around 80 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Nectin-4/PVRL4 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) | 75 | 209 | 857 | 70 | 228 | 777 |
| Standard deviation | 3.67 | 13.16 | 44.56 | 4.13 | 16.41 | 41.18 |
| CV (%) | 4.9% | 7.2%% | 5.2% | 5.9% | 7.2% | 5.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Nectin-4/PVRL4 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 | 75 | 82 | 86 | 80 | 80 | 3.96 | 4.9% |
| Sample 2 | 209 | 190 | 207 | 207 | 203 | 7.69 | 3.7% |
| Sample 3 | 857 | 1006 | 836 | 933 | 908 | 67.1 | 7.3% |
Protein Target Info & Infographic
Gene/Protein Information For NECTIN4 (Source: Uniprot.Org, NCBI)
Gene Name
NECTIN4
Full Name
Nectin-4
Weight
55454 MW
Superfamily
nectin family
Alternative Names
Nectin-4;Ig superfamily receptor LNIR;Nectin cell adhesion molecule 4 ;Poliovirus receptor-related protein 4;Processed poliovirus receptor-related protein 4;NECTIN4 ;LNIR, PRR4, PVRL4; Nectin4|1200017F15Rik, P, Prr4, Pvrl4|nectin cell adhesion molecule 4|nectin-4|ig superfamily receptor LNIR|nectin 4|poliovirus receptor-related 4|poliovirus receptor-related protein 4
*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 NECTIN4, check out the NECTIN4 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 NECTIN4: 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 Nectin-4/PVRL4 ELISA Kit PicoKine® (EK1536)
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No publications found for EK1536
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Customer Reviews
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Customer Q&As
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9 Customer Q&As for Human Nectin-4/PVRL4 ELISA Kit PicoKine®
Question
Q: we need your recommendation regarding the dilution ratio of serum samples for detection of Nectin-4 in Human serum? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2020-02-28
Answer
A: having little idea about 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 Nectin-4 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: 2020-02-28
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2019-08-24
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: 2019-08-24
Question
If the samples you tested were below the detection range for both serum and plasma, what sample type do you think would give values within the actual curve?
Verified Customer
Verified customer
Asked: 2019-05-15
Answer
The samples we tested are serum and plasma samples collected from healthy human. Pathological samples or treated samples might give higer values.
Boster Scientific Support
Answered: 2019-05-15
Question
Q: can I use citrate plasma as samples in Human Nectin-4 Picokine® ELISA Kit (Catalog # EK1536)?
P. Williams
Verified customer
Asked: 2018-07-19
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of Nectin-4 causing degradation of its protein structure. Nectin-4 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 Nectin-4 ELISA, treating samples with a number of 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: 2018-07-19
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
M. Walker
Verified customer
Asked: 2018-06-06
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: 2018-06-06
Question
Q: if the enzyme conjugated Nectin-4 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 Nectin-4 antigen?
T. Hall
Verified customer
Asked: 2018-04-26
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: 2018-04-26
Question
Q: What is the optimal O.D. value for Nectin-4 ELISA kit? I used your Nectin-4 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. is it safe to say these samples contain Nectin-4 even though the O.D. values are not very high?
T. Allen
Verified customer
Asked: 2018-02-06
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. a point of focus should be 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 proportion 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 Nectin-4 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Nectin-4 that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2018-02-06
Question
Q: Can Nectin-4 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Y. Babu
Verified customer
Asked: 2017-12-28
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 check 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: 2017-12-28
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2016-07-03
Answer
A: for 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 can have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2016-07-03
