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Product Info Summary
| SKU: | EK1101 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human Seprase/FAP ELISA Kit PicoKine®
View all Fibroblast Activation Protein alpha/FAP ELISA kits
SKU/Catalog Number
EK1101
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human Seprase/FAP ELISA Kit PicoKine® (96 Tests). Quantitate Human FAP in cell culture supernatants, cell lysates, 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
Human Seprase/FAP ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1101)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: L26-D760
Sensitivity
<10 pg/ml
Assay Range
62.5 pg/ml - 4,000 pg/ml
Standard Dilution Instructions
See datasheet of EK1101 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1101 is reactive to FAP in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK1101 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of Fibroblast Activation Protein alpha/FAP
FAP (Fibroblast Activation Protein, Alpha) also known as FAPA or SEPRASE, is an inducible cell surface glycoprotein that was originally identified in cultured fibroblasts using monoclonal antibody F19. The protein encoded by this gene is a homodimeric integral membrane gelatinase belonging to the serine protease family. The FAP gene is mapped on 2q24.2. FAP is most closely related to DPPIV and they share about 50% of their amino acids. FAP is catalytically active as a 170kD dimer and has dipeptidase and gelatinase activity. Its gelatinase activity requires a glycine in P2 position.FAP-alpha shows 48% amino acid identity with dipeptidyl peptidase IV and 30% identity with DPP4-related protein. Northern blot analysis detected a 2.8-kb FAP-alpha mRNA in fibroblasts. Depletion of FAP-expressing cells, which made up only 2% of all tumor cells in established Lewis lung carcinomas, caused rapid hypoxic necrosis of both cancer and stromal cells in immunogenic tumors by a process involving interferon-gamma and tumor necrosis factor-alpha.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1101-CAP | Anti-Human FAP Pre-coated 96-well strip microplate | 1 |
| EK1101-ST | Human FAP Standard | 2 vials, 10 ng/tube |
| EK1101-DA | Human FAP 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 | 62.5 | 125 | 250 | 500 | 1000 | 2000 | 4000 |
| O.D. | 0.036 | 0.121 | 0.183 | 0.317 | 0.543 | 0.968 | 1.566 | 2.037 |
Data Example Images
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Human Seprase/FAP PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect Fibroblast Activation Protein alpha/FAP, Dilution ratio of 1:100, concentration in serum and plasma is around 60 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human Seprase/FAP 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) | 179 | 783 | 1188 | 189 | 736 | 1293 |
| Standard deviation | 8.59 | 43.84 | 80.78 | 11.52 | 49.31 | 96.97 |
| CV (%) | 4.8% | 5.6% | 6.8% | 6.1% | 6.7% | 7.5% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of Fibroblast Activation Protein alpha/FAP 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 | 179 | 180 | 182 | 195 | 184 | 6.44 | 3.5% |
| Sample 2 | 783 | 741 | 715 | 847 | 771 | 49.88 | 6.4% |
| Sample 3 | 1188 | 1246 | 1205 | 1210 | 1212 | 21.12 | 1.7% |
Protein Target Info & Infographic
Gene/Protein Information For FAP (Source: Uniprot.Org, NCBI)
Gene Name
FAP
Full Name
Prolyl endopeptidase FAP
Weight
87713 MW
Superfamily
peptidase S9B family
Alternative Names
Prolyl endopeptidase FAP ;3.4.21.26 ;170 kDa melanoma membrane-bound gelatinase ;Dipeptidyl peptidase FAP ;3.4.14.5 ;Fibroblast activation protein alpha ;FAPalpha ;Gelatine degradation protease FAP ;3.4.21.- ;Integral membrane serine protease ;Post-proline cleaving enzyme ;Serine integral membrane protease ;SIMP ;Surface-expressed protease ;Seprase ;Antiplasmin-cleaving enzyme FAP, soluble form ;APCE ;3.4.14.5 ;3.4.21.- ;3.4.21.26 ;FAP ; FAP DPPIVA, FAPalpha, SIMP, FAP fibroblast activation protein alpha prolyl endopeptidase FAP|170 kDa melanoma membrane-bound gelatinase|dipeptidyl peptidase FAP|gelatine degradation protease FAP|integral membrane serine protease|post-proline cleaving enzyme|seprase|surface-expressed protease
*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 FAP, check out the FAP 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 FAP: 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 Seprase/FAP ELISA Kit PicoKine® (EK1101)
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Customer Reviews
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Customer Q&As
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8 Customer Q&As for Human Seprase/FAP ELISA Kit PicoKine®
Question
Q: if the enzyme conjugated FAP 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 FAP antigen?
Verified Customer
Verified customer
Asked: 2020-09-06
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: 2020-09-06
Question
Q: how much samples can be assayed in a Picokine® ELISA Kit?
B. Hernandez
Verified customer
Asked: 2020-08-03
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: 2020-08-03
Question
Q: Can FAP ELISA Kits be used with tissue homogenates (or other non-validated sample types)?
C. Gandhi
Verified customer
Asked: 2019-11-07
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 assessd 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: 2019-11-07
Question
Q: can I use citrate plasma as samples in Human FAP Picokine® ELISA Kit (Catalog # EK1101)?
D. Tiwari
Verified customer
Asked: 2019-07-26
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of FAP causing disruption of its protein structure. FAP 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 FAP ELISA, treating samples with different 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: 2019-07-26
Question
Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement FAP using the EK1101 Human FAP Picokine® ELISA Kit?
B. Miller
Verified customer
Asked: 2019-02-26
Answer
A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may affect 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 repetitive freeze-thaw cycles with the stored samples to limit protein degradation.
Boster Scientific Support
Answered: 2019-02-26
Question
Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?
P. Smith
Verified customer
Asked: 2018-10-18
Answer
A: in 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 impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-10-18
Question
Q: which procedure should I follow in order to thaw whole blood sample for FAP ELISA after freezing?
Verified Customer
Verified customer
Asked: 2018-06-01
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 FAP for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.
Boster Scientific Support
Answered: 2018-06-01
Question
Q: how to proceed with the analysis of ELISA data? I measured FAP level in serum.
K. Wright
Verified customer
Asked: 2015-06-11
Answer
A: please read 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: 2015-06-11
