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Product Info Summary
| SKU: | EK1999 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA or citrate) |
Product info
Product Name
Human Nephrin ELISA Kit PicoKine®
SKU/Catalog Number
EK1999
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Nephrin ELISA Kit PicoKine® (96 Tests). Quantitate Human NPHS1 in cell culture supernatants, serum and plasma (heparin, EDTA or citrate) . Sensitivity: 50pg/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 Nephrin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1999)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: Q23-S1055
Sensitivity
<50 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1999 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1999 is reactive to NPHS1 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA or citrate)
Application Guarantee
EK1999 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Nephrin
Nephrin is a protein necessary for the proper functioning of the renal filtration barrier. It is mapped to 19q13.12. This gene encodes a member of the immunoglobulin family of cell adhesion molecules that functions in the glomerular filtration barrier in the kidney. The gene is primarily expressed in renal tissues, and the protein is a type-1 transmembrane protein found at the slit diaphragm of glomerular podocytes. The slit diaphragm is thought to function as an ultrafilter to exclude albumin and other plasma macromolecules in the formation of urine. Mutations in this gene result in Finnish-type congenital nephrosis 1, characterized by severe proteinuria and loss of the slit diaphragm and foot processes.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1999-CAP | Anti-Human NPHS1 Pre-coated 96-well strip microplate | 1 |
| EK1999-ST | Human NPHS1 Standard | 2 vials, 10 ng/tube |
| EK1999-DA | Human NPHS1 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. | 0.055 | 0.091 | 0.172 | 0.242 | 0.417 | 0.683 | 1.221 | 1.826 |
Data Example Images
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Human Nephrin PicoKine ELISA Kit Standard Curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Nephrin, Dilution ratio of 1:1, concentration in serum and plasma is around 200 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Nephrin 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) | 210 | 1142 | 4995 | 287 | 1167 | 49998 |
| Standard deviation | 8.61 | 51.39 | 234.77 | 19.52 | 68.85 | 3049.88 |
| CV (%) | 4.1% | 4.5% | 4.7% | 6.8% | 5.9% | 6.1% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Nephrin 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 | 210 | 216 | 255 | 248 | 232 | 19.52 | 8.4% |
| Sample 2 | 1142 | 1344 | 1347 | 1325 | 1289 | 85.57 | 6.6% |
| Sample 3 | 4995 | 4549 | 4974 | 5064 | 4895 | 202.80 | 4.1% |
Protein Target Info & Infographic
Gene/Protein Information For NPHS1 (Source: Uniprot.Org, NCBI)
Gene Name
NPHS1
Full Name
Nephrin
Weight
134.742kDa
Superfamily
immunoglobulin superfamily
Alternative Names
Nephrin; Renal glomerulus-specific cell adhesion receptor; NPHS1; NPHN NPHS1 CNF, NPHN, nephrin NPHS1 adhesion molecule, nephrin nephrin|NPHS1, nephrin|nephrosis 1, congenital, Finnish type (nephrin)|renal glomerulus-specific cell adhesion receptor|truncated NPHS1
*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 NPHS1, check out the NPHS1 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 NPHS1: 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 Nephrin ELISA Kit PicoKine® (EK1999)
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Customer Reviews
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Customer Q&As
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7 Customer Q&As for Human Nephrin ELISA Kit PicoKine®
Question
Is EK1999 compatible with urine as a specimen?
Verified customer
Asked: 2022-08-11
Answer
Our lab hasn't worked on urine samples using the Human Nephrin ELISA Kit PicoKine® (EK1999). We suggest to run a pilot test to see if the kit could work for the specific sample. In theory, if the expression level of Nephrin in the samples falls in the assay range of the kit, the kit is able to detect the protein in the samples.
Boster Scientific Support
Answered: 2022-08-12
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2020-01-13
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 depends upon the kit used so please refer to each datasheet for details.
Boster Scientific Support
Answered: 2020-01-13
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement Nephrin using the EK1999 Human Nephrin Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-06-01
Answer
A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample on priority basis collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples in order to limit protein degradation.
Boster Scientific Support
Answered: 2019-06-01
Question
Q: What is the optimal O.D. value for Nephrin ELISA kit? I performed your Nephrin 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 Nephrin even though the O.D. values are not very high?
Z. Wang
Verified customer
Asked: 2018-07-25
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. in your example, 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 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 Nephrin ELISA kit to have sensitivity of 50pg/ml, that means the minimum amount of Nephrin that can be declared/interpreted as positive by the above standard is 50pg/ml.
Boster Scientific Support
Answered: 2018-07-25
Question
Q: can you suggest the dilution ratio of serum samples for detection of Nephrin in Human serum? I am trying to measure a multiple analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
M. Perez
Verified customer
Asked: 2018-05-19
Answer
A: without having an understanding 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 Nephrin 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: 2018-05-19
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
Verified Customer
Verified customer
Asked: 2017-02-26
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2017-02-26
Question
Q: Can Nephrin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
Verified Customer
Verified customer
Asked: 2015-01-14
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: 2015-01-14
