TNF alpha ELISA Kit

Produktnummer ABIN1112686

Human TNF alpha ELISA Kit, Colorimetric Assay für die Quantifizierung von Human TNF alpha.

Kurzübersicht für TNF alpha ELISA Kit (ABIN1112686)

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

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 15.6 pg/mL - 1000 pg/mL

Applikation: ELISA

Proben: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Produktdetails TNF alpha ELISA Kit

Untere Nachweisgrenze: 15.6 pg/mL

Verwendungszweck: For quantitative detection of TNFα in Human serum, plasma, tissue lysatesor cell culture supernatants.

Analytische Methode: Quantitative

Sensitivität: < 4 pg/mL

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

Benötigtes Material:

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

Anwendungsinformationen

Applikationshinweise: Optimal working dilution should be determined by the investigator.

Kommentare:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- TNF? polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- TNF? 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 TNF? amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of TNF? can be calculated.
15.6 - 1000 pg/mL (body fluids, lysates, supernates), 7.8 - 500 pg/mL (serum)

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

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.

Aufbereitung der Proben:

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.
Tissue lysate, body fluids or 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) or coat at 4°C overnight. Centrifuge at approximately 1000 × g for 10 min. Analyze the serum immediately or aliquot and store 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konservierungsmittel: Sodium azide, Thimerosal (Merthiolate)

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Store at 2-8°C for 6 months, or at -20°C for 12 months.

Haltbarkeit: 12 months

Details zu TNF alpha

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

Andere Bezeichnung: TNFalpha

Hintergrund: Tumor necrosis factor, formerly known as tumor necrosis factor-alpha or TNF-Į, is a cytokine involved in systemic inflammation and is a member of a group of cytokines that stimulate the acute phase reaction. It was originally identified in Human serum after injection with Mycobacterium bovis strain bacillus Calmette-Guerin (BCG) and endotoxin. TNF is primarily produced as a 212-amino acid-long type II transmembrane protein arranged in stable homotrimers. The human TNF gene maps to chromosome 6p21.3, spans about 3 kilobases and contains 4 exons. The primary role of TNF is in the regulation of immune cells. Dysregulation of TNF production has been implicated in a variety of human diseases, including Alzheimer's disease, cancer, major depression, and inflammatory bowel disease (IBD).

Pathways: NF-kappaB Signalweg, Apoptose, Caspase Kaskade in der Apoptose, TLR Signalweg, 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