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Product Info Summary
| SKU: | EK1487 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and urine. |
Product info
Product Name
Human Afamin ELISA Kit PicoKine®
SKU/Catalog Number
EK1487
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Afamin ELISA Kit PicoKine® (96 Tests). Quantitate Human AFM in cell culture supernatants, serum 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 Afamin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1487)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: L22-N599
Capture/Detection Antibodies
The capture antibody is monoclonal antibody from mouse 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 EK1487 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1487 is reactive to AFM in Human samples
Validated Sample Types
cell culture supernatants, serum and urine.
Application Guarantee
EK1487 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of AFM
Afamin is a protein that in humans is encoded by the AFM gene. This gene is a member of the albumin gene family, which is comprised of four genes that localize to chromosome 4 in a tandem arrangement. These four genes encode structurally-related serum transport proteins that are known to be evolutionarily related. The protein encoded by this gene is regulated developmentally, expressed in the liver and secreted into the bloodstream.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1487-CAP | Anti-Human AFM Pre-coated 96-well strip microplate | 1 |
| EK1487-ST | Human AFM Standard | 2 vials, 10 ng/tube |
| EK1487-DA | Human AFM 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. | 0.068 | 0.093 | 0.121 | 0.193 | 0.354 | 0.715 | 1.559 | 2.457 |
Data Example Images
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Human Afamin PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect AFM, Dilution ratio of 1:100000, concentration in serum is around 25 ug/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Afamin 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) | 221 | 658 | 3779 | 249 | 672 | 4128 |
| Standard deviation | 10.83 | 33.56 | 219.18 | 17.68 | 51.07 | 321.98 |
| CV (%) | 4.9% | 5.1% | 5.8% | 7.1% | 7.6% | 7.8% |
Specific Publications For Human Afamin ELISA Kit PicoKine® (EK1487)
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Customer Reviews
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Customer Q&As
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8 Customer Q&As for Human Afamin ELISA Kit PicoKine®
Question
Q: can you suggest the dilution ratio of serum samples for detection of Afamin in Human urine? I am trying to measure a a number of 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.
Verified Customer
Verified customer
Asked: 2020-10-17
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 Afamin 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: 2020-10-17
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2020-01-28
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: 2020-01-28
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification Afamin using the EK1487 Human Afamin Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-11-07
Answer
A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may change 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. avoid repeat freeze-thaw cycles with the stored samples to prevent protein degradation.
Boster Scientific Support
Answered: 2019-11-07
Question
What specific cell line was used when validating EK1487 kit for cell culture supernant samples?
Verified customer
Asked: 2019-05-07
Answer
We tested PBMC cell line for the Human Afamin ELISA Kit PicoKine™ (EK1487).
Boster Scientific Support
Answered: 2019-05-07
Question
Q: which procedure should I follow in order to thaw whole blood sample for Afamin ELISA after freezing?
K. Scott
Verified customer
Asked: 2019-03-17
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 Afamin for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.
Boster Scientific Support
Answered: 2019-03-17
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
Verified Customer
Verified customer
Asked: 2018-08-19
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: 2018-08-19
Question
Q: if the enzyme conjugated Afamin antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Afamin antigen?
Verified Customer
Verified customer
Asked: 2018-07-30
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: 2018-07-30
Question
Q: Can Afamin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
W. Harris
Verified customer
Asked: 2017-11-13
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 suitable 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 view 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: 2017-11-13
