Human Seprase/FAP PicoKine™ ELISA Kit
|Size||96wells/kit, with removable strips.|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA).|
|Product Name||Human Seprase/FAP PicoKine™ ELISA Kit|
|Storage & Handling||Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)|
|Size||96wells/kit, with removable strips.|
|Description||Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Seprase/FAP. 96wells/kit, with removable strips.|
|Cite This Product||Human Seprase/FAP PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1101)|
|Sample Type||cell culture supernates, cell lysates, serum and plasma (heparin, EDTA)..
Anticoagulant(s): heparin or EDTA
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility.
|Immunogen||Expression system for standard: NSO; Immunogen sequence: L26-D760|
|Cross Reactivity||There is no detectable cross-reactivity with other relevant proteins.|
|Antibody Clonalities||Capture Antibody | Detection Antibody:
monoclonal antibody from mouse|polyclonal antibody from goat
|EK1101-CAP||96-well plate precoated with anti-Human FAP antibody||1|
|EK1101-ST||lyophilized recombinant Human FAP standard||10ng/tube|
|EK1101-DA||biotinylated anti-Human FAP antibody||130ul|
|AR1106-1||sample diluent buffer||30ml|
|AR1106-2||antibody diluent buffer||12ml|
|AR1106-3||ABC diluent buffer||12ml|
|AR1104||TMB color developing agent||10ml|
|AR1105||TMB stop solution||10ml|
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human Seprase/FAP PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
Intra/Inter Assay Precision
|Intra-Assay Precision||Inter-Assay Precision|
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
|Lots||Lot1 (pg/ml)||Lot2 (pg/ml)||Lot3 (pg/ml)||Lot4 (pg/ml)||Mean (pg/ml)||Standard Deviation||CV (%)|
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Prolyl endopeptidase FAP|
|Tissue Specificity||Expressed in adipose tissue. Expressed in the dermal fibroblasts in the fetal skin. Expressed in the granulation tissue of healing wounds and on reactive stromal fibroblast in epithelial cancers. Expressed in activated fibroblast-like synoviocytes from inflamed synovial tissues. Expressed in activated hepatic stellate cells (HSC) and myofibroblasts from cirrhotic liver, but not detected in normal liver. Expressed in glioma cells (at protein level). Expressed in glioblastomas and glioma cells. Isoform 1 and isoform 2 are expressed in melanoma, carcinoma and fibroblast cell lines. .|
|Alternative Names||Prolyl endopeptidase FAP ;188.8.131.52 ;170 kDa melanoma membrane-bound gelatinase ;Dipeptidyl peptidase FAP ;184.108.40.206 ;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 ;220.127.116.11 ;3.4.21.- ;18.104.22.168 ;FAP ;|
|Subcellular Localization||Prolyl endopeptidase FAP: Cell surface . Cell membrane ; Single- pass type II membrane protein . Cell projection, lamellipodium membrane ; Single-pass type II membrane protein . Cell projection, invadopodium membrane ; Single-pass type II membrane protein . Cell projection, ruffle membrane ; Single-pass type II membrane protein . Membrane ; Single-pass type II membrane protein . Localized on cell surface with lamellipodia and invadopodia membranes and on shed vesicles. Colocalized with DPP4 at invadopodia and lamellipodia membranes of migratory activated endothelial cells in collagenous matrix. Colocalized with DPP4 on endothelial cells of capillary-like microvessels but not large vessels within invasive breast ductal carcinoma. Anchored and enriched preferentially by integrin alpha-3/beta-1 at invadopodia, plasma membrane protrusions that correspond to sites of cell invasion, in a collagen-dependent manner. Localized at plasma and ruffle membranes in a collagen-independent manner. Colocalized with PLAUR preferentially at the cell surface of invadopodia membranes in a cytoskeleton-, integrin- and vitronectin-dependent manner. Concentrated at invadopodia membranes, specialized protrusions of the ventral plasma membrane in a fibrobectin-dependent manner. Colocalizes with extracellular components (ECM), such as collagen fibers and fibronectin. .|
|Molecular Weight||87713 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Cell surface glycoprotein serine protease that participates in extracellular matrix degradation and involved in many cellular processes including tissue remodeling, fibrosis, wound healing, inflammation and tumor growth. Both plasma membrane and soluble forms exhibit post-proline cleaving endopeptidase activity, with a marked preference for Ala/Ser-Gly-Pro-Ser/Asn/Ala consensus sequences, on substrate such as alpha-2-antiplasmin SERPINF2 and SPRY2 (PubMed:14751930, PubMed:16223769, PubMed:16480718, PubMed:16410248, PubMed:17381073, PubMed:18095711, PubMed:21288888, PubMed:24371721). Degrade also gelatin, heat-denatured type I collagen, but not native collagen type I and IV, vibronectin, tenascin, laminin, fibronectin, fibrin or casein (PubMed:9065413, PubMed:2172980, PubMed:7923219, PubMed:10347120, PubMed:10455171, PubMed:12376466, PubMed:16223769, PubMed:16651416, PubMed:18095711). Have also dipeptidyl peptidase activity, exhibiting the ability to hydrolyze the prolyl bond two residues from the N-terminus of synthetic dipeptide substrates provided that the penultimate residue is proline, with a preference for Ala-Pro, Ile-Pro, Gly-Pro, Arg-Pro and Pro-Pro (PubMed:10347120, PubMed:10593948, PubMed:16175601, PubMed:16223769, PubMed:16651416, PubMed:16410248, PubMed:17381073, PubMed:21314817, PubMed:24371721, PubMed:24717288). Natural neuropeptide hormones for dipeptidyl peptidase are the neuropeptide Y (NPY), peptide YY (PYY), substance P (TAC1) and brain natriuretic peptide 32 (NPPB) (PubMed:21314817). The plasma membrane form, in association with either DPP4, PLAUR or integrins, is involved in the pericellular proteolysis of the extracellular matrix (ECM), and hence promotes cell adhesion, migration and invasion through the ECM. Plays a role in tissue remodeling during development and wound healing. Participates in the cell invasiveness towards the ECM in malignant melanoma cancers. Enhances tumor growth progression by increasing angiogenesis, collagen fiber degradation and apoptosis and by reducing antitumor response of the immune system. Promotes glioma cell invasion through the brain parenchyma by degrading the proteoglycan brevican. Acts as a tumor suppressor in melanocytic cells through regulation of cell proliferation and survival in a serine protease activity-independent manner. .|
|Background||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.|
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• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
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