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Product Info Summary
| SKU: | EK1661 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Mouse |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, serum and plasma (heparin, EDTA). |
Product info
Product Name
Mouse AFP ELISA Kit PicoKine®
View all alpha-Fetoprotein/AFP ELISA kits
SKU/Catalog Number
EK1661
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Mouse AFP ELISA Kit PicoKine® (96 Tests). Quantitate Mouse Afp in cell culture supernatants, serum and plasma (heparin, EDTA). 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
Mouse AFP ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1661)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: K19-V605
Sensitivity
<10 pg/ml
Assay Range
312 pg/ml - 20,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1661 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1661 is reactive to Afp in Mouse samples
Validated Sample Types
cell culture supernatants, serum and plasma (heparin, EDTA).
Application Guarantee
EK1661 is guaranteed for ELISA in Mouse by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of alpha-Fetoprotein/AFP
alpha-Fetoprotein (AFP), is albuminoid superfamily protein that is synthesized in the fetus primarily by the liver, yolk sac, and tissues of gastrointestinal origin. This gene is mapped to 5 E1; 5 44.77 cM. AFP acts as a carrier protein for steroids, bilirubin, fatty acids, retinoids, and flavonoids. In addition, it can exert immunosuppressive activity, regulate cell proliferation and apoptosis, initiate intracellular signaling, and contribute to cell invasion. Altered levels of both fetal and maternal AFP have been associated with hypothyroidism, autoimmune disorders, and heart defects. Low maternal serum AFP levels are associated with a higher incidence of Down syndrome, whereas higher levels are associated with spina bifida and anencephaly. Elevated AFP levels are also coincident with liver, stomach, and germ cell cancers.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1661-CAP | Anti-Mouse Afp Pre-coated 96-well strip microplate | 1 |
| EK1661-ST | Mouse Afp Standard | 2 vials, 20 ng/tube |
| EK1661-DA | Mouse Afp 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 | 312 | 625 | 1250 | 2500 | 5000 | 10000 | 20000 |
| O.D. | 0.006 | 0.065 | 0.134 | 0.269 | 0.466 | 0.924 | 1.552 | 2.066 |
Data Example Images
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Mouse AFP PicoKine ELISA Kit Standard Curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect alpha-Fetoprotein/AFP, Dilution ratio of 1:10, concentration in serum and plasma is 10-80 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Mouse AFP 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) | 787 | 2378 | 9095 | 804 | 2193 | 8576 |
| Standard deviation | 44.07 | 149.81 | 436.56 | 49.84 | 144.73 | 471.68 |
| CV (%) | 5.6% | 6.3% | 4.8% | 6.2% | 6.6% | 5.5% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of alpha-Fetoprotein/AFP 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 | 787 | 751 | 817 | 732 | 771 | 32.75 | 4.2% |
| Sample 2 | 2378 | 2440 | 2261 | 2414 | 2373 | 68.44 | 2.8% |
| Sample 3 | 9095 | 10188 | 9364 | 8739 | 9346 | 534.02 | 5.7% |
Protein Target Info & Infographic
Gene/Protein Information For Afp (Source: Uniprot.Org, NCBI)
Gene Name
Afp
Full Name
Alpha-fetoprotein
Weight
68.678kDa
Superfamily
ALB/AFP/VDB family
Alternative Names
Alpha-fetoprotein; Alpha-1-fetoprotein; Alpha-fetoglobulin; Afp AFP AFPD, FETA, HPAFP alpha fetoprotein alpha-fetoprotein|alpha-1-fetoprotein|alpha-fetoglobulin
*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 Afp, check out the Afp 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 Afp: 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 Mouse AFP ELISA Kit PicoKine® (EK1661)
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Customer Q&As
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10 Customer Q&As for Mouse AFP ELISA Kit PicoKine®
Question
Q: if the enzyme conjugated alpha-Fetoprotein 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 alpha-Fetoprotein antigen?
B. Anand
Verified customer
Asked: 2020-09-21
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: 2020-09-21
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
K. Phillips
Verified customer
Asked: 2020-08-31
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer may affect immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2020-08-31
Question
Q: is there any online tool I can use to streamline the data analysis for my ELISA results?
Verified Customer
Verified customer
Asked: 2020-07-21
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: 2020-07-21
Question
Q: can I use citrate plasma as samples in Mouse alpha-Fetoprotein Picokine® ELISA Kit (Catalog # EK1661)?
Verified Customer
Verified customer
Asked: 2020-02-28
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of alpha-Fetoprotein causing disruption of its protein structure. alpha-Fetoprotein may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the alpha-Fetoprotein 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: 2020-02-28
Question
Q: we need your recommendation regarding the dilution ratio of serum samples for detection of alpha-Fetoprotein in Mouse 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: 2019-06-03
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 alpha-Fetoprotein 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: 2019-06-03
Question
Q: Are Boster Bio recombinant proteins and antibodies sterile?
M. Hill
Verified customer
Asked: 2019-05-17
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: 2019-05-17
Question
Q: how do I analyze ELISA data? I measured alpha-Fetoprotein level in serum.
Verified Customer
Verified customer
Asked: 2019-03-09
Answer
A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2019-03-09
Question
Q: What is the optimal O.D. value for alpha-Fetoprotein ELISA kit? I performed your alpha-Fetoprotein 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 alpha-Fetoprotein even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2019-01-30
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. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. regarding 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 alpha-Fetoprotein ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of alpha-Fetoprotein that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2019-01-30
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2018-08-10
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: 2018-08-10
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring alpha-Fetoprotein using the EK1661 Mouse alpha-Fetoprotein Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2017-07-26
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 immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples in order to limit protein degradation.
Boster Scientific Support
Answered: 2017-07-26
