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Product Info Summary
| SKU: | EK0349 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA, citrate). |
Product info
Product Name
Human Fibronectin ELISA Kit PicoKine®
SKU/Catalog Number
EK0349
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Fibronectin ELISA Kit PicoKine® (96 Tests). Quantitate Human FN1 in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). 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 Fibronectin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0349)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Target protein isolated from human plasma
Capture/Detection Antibodies
The capture antibody is polyclonal antibody from rabbit and the detection antibody is polyclonal antibody from goat.
Sensitivity
<10 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0349 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0349 is reactive to FN1 in Human samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA, citrate).
Application Guarantee
EK0349 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of FN1
Fibronectin (FN) also known as LETS, is identified on the surfFN of fibroblasts by labeling with radioactive compounds or specific antibodies. Fibronectin is a 430,000-dalton dimeric glycoprotein that exists in 2 forms, termed cellular and plasma fibronectin. Cellular and plasma fibronectins are heterodimers consisting of similar but not identical polypeptides. These two forms of FN differ in biologic activity. Fibronectins bind cell surfFNs and various compounds including collagen, fibrin, heparin, DNA, and actin. Because fibronectin stimulates endocytosis in several systems and promotes the clearance of particulate material from the circulation, it could function in the clearance of C1q-coated material such as immune complexes or cellular debris. Fibronectins are involved in cell adhesion, cell motility, opsonization, would healing, and maintenance of cell shape. LETS, encoded on chromosome 8, is responsible for the LETS protein expression in humans. Because LETS has been implicated in tumorigenicity and cellular transformation, it is of interest that rearrangement or modifications in the number of chromosome 8 have been associated with certain forms of cancer. The standard used in this kit is isolated from human plasma with the molecular mass of 200-250KDa.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0349-CAP | Anti-Human FN1 Pre-coated 96-well strip microplate | 1 |
| EK0349-ST | Human FN1 Standard | 2 vials, 10 ng/tube |
| EK0349-DA | Human FN1 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. | 0.021 | 0.068 | 0.112 | 0.197 | 0.336 | 0.605 | 1.115 | 1.83 |
Data Example Images
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Human Fibronectin PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect FN1, Dilution ratio of 1:10000, concentration in serum and plasma is around 15 ug/ml..
Some articles we found to cite concentrations of FN1 in samples: 25732262, 25551962 (Pubmed IDs).
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Fibronectin 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) | 312 | 1118 | 5412 | 341 | 1105 | 4997 |
| Standard deviation | 14.35 | 65.76 | 308.48 | 19.43 | 76.24 | 349.79 |
| CV (%) | 4.6% | 5.7% | 5.7% | 5.7% | 6.9% | 7% |
Specific Publications For Human Fibronectin ELISA Kit PicoKine® (EK0349)
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Customer Reviews
0 Reviews For Human Fibronectin ELISA Kit PicoKine®
Customer Q&As
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6 Customer Q&As for Human Fibronectin ELISA Kit PicoKine®
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of Fibronectin in Human serum? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.
J. Perez
Verified customer
Asked: 2020-10-12
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 Fibronectin 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: 2020-10-12
Question
Q: can you tell me how to prepare cell lysates prepared for use in Picokine® ELISA kits?
B. Bhasin
Verified customer
Asked: 2019-10-24
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 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: 2019-10-24
Question
Do you know if EK0349 might also work with urine samples?
Verified customer
Asked: 2019-05-28
Answer
For Human Fibronectin PicoKine™ ELISA Kit EK0349, we have not tested this kit for urine samples. In theory, the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. For sample types that we have not tested for, we suggest to run pilot experiments and decide the optimal sample dilution.
Boster Scientific Support
Answered: 2019-05-28
Question
Q: Can Fibronectin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
C. Patel
Verified customer
Asked: 2018-06-20
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 needed. 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: 2018-06-20
Question
Q: What is the optimal O.D. value for Fibronectin ELISA kit? I performed your Fibronectin 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 Fibronectin even though the O.D. values are not very high?
V. Nelson
Verified customer
Asked: 2018-04-09
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 the example mentioned, 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. 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 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 Fibronectin ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Fibronectin that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2018-04-09
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring Fibronectin using the EK0349 Human Fibronectin Picokine® ELISA Kit?
K. Moore
Verified customer
Asked: 2016-04-08
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may alter the stabilityof samples. It is recommend to assay sample right after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples to prevent protein degradation.
Boster Scientific Support
Answered: 2016-04-08
