FAS ELISA Kit

Catalog No. ABIN1112600

This Colorimetric ELISA kit is designed for the quantitative measurement of Mouse FAS.

Quick Overview for FAS ELISA Kit (ABIN1112600)

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

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2 pg/mL - 2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Product Details FAS ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: For quantitative detection of FAS in mouse serum, plasma, tissue lysates,cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 3 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Mouse FAS antibody 2. Lyophilized Mouse FAS standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Mouse FAS antibody (Concentrated): 130 µl.

Material not included:

1. 37 °C incubator
2. Microplate reader (wavelength: 450 nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5 mL of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1 L

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-FAS polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-FAS polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the FAS amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of FAS can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37 °C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Body fluids, tissue lysates and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 10 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin, EDTA or citrate as the anticoagulant. Centrifuge for 10 min at 1000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Storage: 4 °C,-20 °C

Storage Comment: Store at 2-8°C for 6 months, or at -20°C for 12 months.

Expiry Date: 12 months

Target Details for FAS

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

Alternative Name: FAS / APO-1 / CD95

Background: Fas, also known as CD95, APO-1 and TNFRSF6, is a member of the TNF-receptor superfamily. This receptor contains a death domain. It has been shown to play a central role in the physiological regulation of programmed cell death, and it has been implicated in the pathogenesis of various malignancies and diseases of the immune system. The interaction of Fas with its ligand allows the formation of a death-inducing signaling complex that includes Fas-associated death domain protein (FADD), caspase 8, and caspase 10. The autoproteolytic processing of the caspases in the complex triggers a downstream caspase cascade and leads to apoptosis. This receptor has been also shown to activate NF-kappaB, MAPK3/ERK1, and MAPK8/JNK, and it is found to be involved in transducing the proliferating signals in normal diploid fibroblast and T cells.

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