TNF alpha IQ-ELISA Kit

Catalog No. ABIN6244368

Product Details TNF alpha IQ-ELISA Kit

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

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Detection Range: 0.384 pg/mL - 6000 pg/mL

Minimum Detection Limit: 0.384 pg/mL

Application: ELISA

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

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Semi-Quantitative

Characteristics: 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.

Components:

• 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

Material not included:

• 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

Application Details

Application Notes: 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.

Comment:

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

Plate: Pre-coated

Protocol:

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

Reagent Preparation:

Recommended Dilution for serum and plasma samples2-10 fold

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: 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.

Expiry Date: 6 months

Target Details

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

Alternative Name: TNF alpha (TNF alpha Products)

Synonyms: DIF IQ-ELISA Kit, TNF-alpha IQ-ELISA Kit, TNFA IQ-ELISA Kit, TNFSF2 IQ-ELISA Kit, RATTNF IQ-ELISA Kit, Tnfa IQ-ELISA Kit, tnf IQ-ELISA Kit, TNF-a IQ-ELISA Kit, TNFalpha IQ-ELISA Kit, Tnfsf1a IQ-ELISA Kit, TNFa IQ-ELISA Kit, cTNF IQ-ELISA Kit, Tnf-alpha IQ-ELISA Kit, tnfa-like IQ-ELISA Kit, TNF-ALPHA IQ-ELISA Kit, dif IQ-ELISA Kit, tnfa IQ-ELISA Kit, xtnf IQ-ELISA Kit, tnfsf2 IQ-ELISA Kit, tnf-alpha IQ-ELISA Kit, Cachectin IQ-ELISA Kit, tumor necrosis factor IQ-ELISA Kit, tumor necrosis factor b (TNF superfamily, member 2) IQ-ELISA Kit, tumor necrosis factor alpha IQ-ELISA Kit, tumor necrosis factor a (TNF superfamily, member 2) IQ-ELISA Kit, TNF IQ-ELISA Kit, Tnf IQ-ELISA Kit, tnf IQ-ELISA Kit, tnfb IQ-ELISA Kit, tnf-alpha IQ-ELISA Kit, LOC103694380 IQ-ELISA Kit, tnfa IQ-ELISA Kit

Gene ID: 7124

UniProt: P01375

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