TNF alpha Kit IQ-ELISA

N° du produit ABIN6244368

Ce kit ELISA qPCR est conçu pour la mesure quantitative de Humain TNF alpha.

Aperçu rapide pour TNF alpha Kit IQ-ELISA (ABIN6244368)

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

Reactivité: Humain

Méthode de détection: qPCR

Type de méthode: Sandwich ELISA

Gamme de detection: 0.384 pg/mL - 6000 pg/mL

Application: ELISA

Type d'échantillon: Cell Culture Supernatant, Plasma, Serum

Détail du produit TNF alpha Kit IQ-ELISA

Seuil minimal de détection: 0.384 pg/mL

Fonction: Human Immunoquantitative (PCR-Based) TNF alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.

Analytical Method: Semi-Quantitative

Attributs du produit: IQELISAs employ specific capture antibodies coated on a 96-well PCR plate. Standards and samples are pipetted into the wells; the target protein in the standards and samples binds to the immobilized antibody. The wells are washed and the detection affinity reagent is added to the wells where it binds to any captured antigen. The wells are washed, and primers and PCR master mix are added to each well. The plate is placed into a real time PCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of affinity detection reagent that bound to captured antigen in each well.

Ingrédients:

• TNF alpha Microplate (Item A): 96 well PCR plate coated with anti-Human TNF alpha
• Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
• Standards (Item C): 2 vials of recombinant Human TNF alpha
• Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
• Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
• Detection Affinity Reagent for TNF alpha (Item F): 2 vials of a 4x concentrated solution of anti-Human TNF alpha affinity reagent
• IQELISA Detection Reagent (Item G): 1.4ml of a 10x concentrated stock
• Primer Solution (Item I): 1.7 ml vial
• PCR Master Mix (Item J): 1.2 ml vial
• PCR Preparation buffer (Item K): 1ml vial of 10x concentrated buffer
• Final Wash Buffer (Item L): 10ml vial of 10x concentrated buffer

Matériel non inclus:

• Real-time PCR instrument, Bio-Rad recommended
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for data analysis
• Tubes to prepare standard or sample dilutions
• Heating block or water bath capable of 80^°C

Information d'application

Indications d'application: The Immuno-Quantitative ELISA (IQELISA) kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of only 1/10th the sample volume while also providing 10x more sensitivity than a traditional ELISA.

Commentaires:

Features:
- Excellent for follow up/confirmation of antibody array data
- Pre-coated 96 well plate
- Strip plate format minimizes waste
- Complete kit includes all necessary reagents
- 10x more sensitive than ELISA
- Reproducible and reliable
- Optimized for serum, plasma, urine and cell culture medium
- 1/10th the sample volume of an ELISA

Research Applications:
- Detection of quantitative protein levels in biological fluids
- Validation of antibody array results
- Validation of biomarker discovery studies

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed
2. Add 25 μL standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4^°C
3. Add 25 μL detection affinity reagent to each well. Incubate 1 hour at room temperature
4. Add 25μL of IQELISA Detection Reagent to each well. Incubate 1 hour
5. Add 15μL Primer solution 10μL of PCR master mix to each well
6. Run real-time PCR

Préparation des réactifs:

Recommended Dilution for serum and plasma samples2-10 fold

Restrictions: For Research Use only

Stockage

Stock: -20 °C

Stockage commentaire: May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.

Date de péremption: 6 months

Détail du antigène

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

Autre désignation: TNF alpha

ID gène: 7124

UniProt: P01375

Pathways: Signalisation NF-kappaB, Apoptose, Caspase Cascade in Apoptosis, Signalisation TLR, 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