TWEAK ELISA Kit

Catalog No. ABIN1889351

Product Details TWEAK ELISA Kit

Target: TWEAK (TNFSF12) (Tumor Necrosis Factor (Ligand) Superfamily, Member 12 (TNFSF12))

Binding Specificity: AA 94-249

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6-1000 pg/mL

Minimum Detection Limit: 15.6 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse TNFSF12/TWEAK

Brand: PicoKine™

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

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: S94-H249

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

Sensitivity: <10pg/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: E.coli
Immunogen sequence: S94-H249

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:

Tissue Specificity: Widely expressed.

Plate: Pre-coated

Protocol: mouse TNFSF12 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for TNFSF12 has been precoated onto 96-well plates. Standards(E.coli, S94-H249) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TNFSF12 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 mouse TNFSF12 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL mouse TNFSF12 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 mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well.. See "Sample Dilution Guideline" above for details. It is recommended that each mouse TNFSF12 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 84, Standard deviation: 3.53, CV(%): 4.2
• Sample 2: n=16, Mean(pg/ml): 210, Standard deviation: 7.35, CV(%): 3.5
• Sample 3: n=16, Mean(pg/ml): 537, Standard deviation: 27.4, CV(%): 5.1,
• Sample 1: n=24, Mean(pg/ml): 106, Standard deviation: 5.62, CV(%): 5.3
• Sample 2: n=24, Mean(pg/ml): 324, Standard deviation: 14.91, CV(%): 4.6
• Sample 3: n=24, Mean(pg/ml): 638, Standard deviation: 38.28, CV(%): 6

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

Target Details for TWEAK

Target: TWEAK (TNFSF12) (Tumor Necrosis Factor (Ligand) Superfamily, Member 12 (TNFSF12))

Alternative Name: TNFSF12 (TNFSF12 Products)

Background:

Protein Function: Binds to FN14 and possibly also to TNRFSF12/APO3. Weak inducer of apoptosis in some cell types. Mediates NF-kappa-B activation. Promotes angiogenesis and the proliferation of endothelial cells. Also involved in induction of inflammatory cytokines. Promotes IL8 secretion (By similarity). .

Background: Tumor necrosis factor ligand superfamily member 12 also known as TNF-related weak inducer of apoptosis(TWEAK) is a protein that in humans is encoded by the TNFSF12 gene. The protein encoded by this gene is a cytokine that belongs to the tumor necrosis factor(TNF) ligand family. This protein is a ligand for the FN14/TWEAKR receptor. This cytokine has overlapping signaling functions with TNF, but displays a much wider tissue distribution. This cytokine can induce apoptosis via multiple pathways of cell death in a cell type-specific manner. This cytokine is also found to promote proliferation and migration of endothelial cells, and thus acts as a regulator ofangiogenesis. The TNFSF12 gene lies 878 bp upstream of the putative transcriptional start site of the TNFSF13 gene on chromosome 17p13.1.

Synonyms: Tumor necrosis factor ligand superfamily member 12,TNF-related weak inducer of apoptosis,TWEAK,Tumor necrosis factor ligand superfamily member 12, membrane form,Tumor necrosis factor ligand superfamily member 12, secreted form,Tnfsf12,

Full Gene Name: Tumor necrosis factor ligand superfamily member 12

Cellular Localisation: Cell membrane, Single-pass type II membrane protein.

Gene ID: 21944

UniProt: O54907

Pathways: Apoptosis