FAS ELISA Kit

Catalog No. ABIN411269

This Colorimetric ELISA kit is designed for the quantitative measurement of Human FAS. There is 1 publication available.

Quick Overview for FAS ELISA Kit (ABIN411269)

Target: FAS (TNF Receptor Superfamily, Member 6 (FAS))

Binding Specificity: AA 17-173

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Plasma (citrate)

Product Details FAS ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human Soluble FAS

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: R17-N173

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <3pg/mL

Material not included: 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

Immunogen: Expression system for standard: NSO
Immunogen sequence: R17-N173

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Contains 1 death domain.

Tissue Specificity: Isoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6. .

Plate: Pre-coated

Protocol: human FAS ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for FAS has been precoated onto 96-well plates. Standards(NSO, R17-N173) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for FAS is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human FAS amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL human FAS standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernatants, serum or plasma(heparin, EDTA, citrate) to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each human FAS standard solution and each sample is measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 138, Standard deviation: 5.80, CV(%): 4.2
• Sample 2: n=16, Mean(pg/ml): 828, Standard deviation: 25.67, CV(%): 3.1
• Sample 3: n=16, Mean(pg/ml): 1507, Standard deviation: 70.83, CV(%): 4.7,
• Sample 1: n=24, Mean(pg/ml): 146, Standard deviation: 10.95, CV(%): 7.5
• Sample 2: n=24, Mean(pg/ml): 839, Standard deviation: 45.31, CV(%): 5.4
• Sample 3: n=24, Mean(pg/ml): 1512, Standard deviation: 83.16, CV(%): 5.5

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for FAS ELISA Kit (ABIN411269)

Liu, Shan, Dong, Liu, Ma, Liu: "Combined early fluid resuscitation and hydrogen inhalation attenuates lung and intestine injury." in: World journal of gastroenterology, Vol. 19, Issue 4, pp. 492-502, (2013) (PubMed).

Target Details for FAS

Target: FAS (TNF Receptor Superfamily, Member 6 (FAS))

Alternative Name: FAS

Background:

Protein Function: Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death- inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS- mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro). .

Background: Fas, also known as APO-1, CD95 and TNFRSF6, is a member of the nerve growth factor(NGF)/tumour necrosis factor(TNF) receptor superfamily and mediates apoptosis. The nucleotide sequence of the cDNAs reveales that the molecule coding for the Fas antigen determinant is a 319 amino acid polypeptide with a single transmembrane domain. The extracellular domain is rich in cysteine residue, and shows a similarity to that of human tumor necrosis factor receptors, human nerve growth factor receptor, and human B cell antigen CD40. The APO-1 antigen as defined by the mouse monoclonal antibody anti-APO-1 is previously found to be expressed on the cell surface of activated human T and B lymphocytes and a variety of malignant human lymphoid cell lines. The APO-1 antigen is found to be a membrane glycoprotein of 48- kDa. Fas antigen is expressed and functional on papillary thyroid cancer cells and this may have potential therapeutic significance. Fas can play a role as an inducer of both neurite growth in vitro and accelerates recovery after nerve injury in vivo. The FAS and FASL triggered apoptosis pathway plays an important role in human carcinogenesis.

Synonyms: Tumor necrosis factor receptor superfamily member 6,Apo-1 antigen,Apoptosis-mediating surface antigen FAS,FASLG receptor,CD95,FAS,APT1, FAS1, TNFRSF6,

Full Gene Name: Tumor necrosis factor receptor superfamily member 6

Cellular Localisation: Isoform 1: Cell membrane, Single-pass type I membrane protein.

Gene ID: 355

UniProt: P25445

Pathways: p53 Signaling, Apoptosis, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity