TNF alpha ELISA Kit

Catalog No. ABIN8056000

The Rat TNF alpha ELISA Kit (ABIN8056000) is a Colorimetric ELISA Kit designed to quantify Rat TNF alpha.

Quick Overview for TNF alpha ELISA Kit (ABIN8056000)

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Binding Specificity: AA 80-235

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 1.56 pg/mL - 100 pg/mL

Application: ELISA

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

Grade: High Sensitivity

Product Details TNF alpha ELISA Kit

Minimum Detection Limit: 1.56 pg/mL

Purpose: High Sensitivity Sandwich Rat TNF Alpha/Tumor necrosis factor High Sensitivity ELISA Kit to quantitate Rat Tnf in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: L80-L235

Capture antibody and Detection antibody: monoclonal antibody from mouse|polyclonal antibody from goat

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

Sensitivity: 0.006 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: 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. 500 mL graduated cylinders. Test tubes for dilution.

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.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target 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 target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 2.0%, 3.8%, 5.0%
Inter-Assay Precision (CV%): 4.9%, 4.7%, 4.9%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: 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.)

Expiry Date: 12 months

Target Details for TNF alpha

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Alternative Name: TNF

Background:

Background: Tumor necrosis factor-alpha (TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor (TNF) and Lymphotoxin (TNF-beta) are cytotoxic proteins which have similar biological activities and share 30 % amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6. The standard product used in this kit is recombinant rat TNFalpha, consisting of 157 amino acids with the molecular mass of 17KDa.

Gene Full Name: tumor necrosis factor

Gene ID: 24835

UniProt: P16599

Pathways: NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome