Interferon gamma IQ-ELISA Kit

Catalog No. ABIN5680034

Product Details Interferon gamma IQ-ELISA Kit

Target: Interferon gamma (IFNG)

Reactivity: Human

Detection Method: qPCR

Method Type: Sandwich ELISA

Application: ELISA

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

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Semi-Quantitative

Sensitivity: 3 pg/mL

Characteristics: The Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA) is an innovative assay 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 lower sample volumes while also providing more sensitivity. The Human IFN-Gamma IQELISA kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of human IFN-Gamma in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for human IFN-Gamma coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and IFN-Gamma present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection affinity molecule is added to the plates. After washing away unbound detection affinity molecule, primers and a PCR master mix are added to the wells and data is collected using qPCR. Ct values obtained from the qPCR are then used to calculate the amount of antigen contained in each sample, where lower Ct values indicate a higher concentration of antigen.

Components:

• IFN-Gamma Microplate (Item A): 96 well PCR plate coated with anti-human IFN-Gamma
• Wash Buffer I Concentrate (20x) (Item B): 20x concentrated solution
• Standards (Item C): recombinant human IFN-Gamma
• Assay Diluent A (Item D): diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
• Assay Diluent B (Item E): 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
• Detection Affinity Reagent for IFN-Gamma (Item F): 4x concentrated solution of anti-human IFN-Gamma affinity reagent
• Primer Solution (Item G)
• PCR Master Mix (Item H)
• PCR Preparation buffer (Item I): 10x concentrated buffer
• Final Wash Buffer (Item J): 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 15μL Primer solution to each well
5. Add 10μL of PCR Master Mix to each well
6. Run real-time PCR

Sample Preparation:

Recommended Dilution (serum & plasma): 3 - 50 fold

Assay Precision: Serum spike tests show recovery is 94.3% with a range of 88.4 to 102.1%

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C,-80 °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

Expiry Date: 6 months

Target Details

Target: Interferon gamma (IFNG)

Alternative Name: IGN-Gamma (IFNG Products)

Synonyms: IFG IQ-ELISA Kit, IFI IQ-ELISA Kit, IFN-g IQ-ELISA Kit, Ifg IQ-ELISA Kit, IFNG2 IQ-ELISA Kit, IFN-gamma IQ-ELISA Kit, IFN-G IQ-ELISA Kit, IFNG IQ-ELISA Kit, IFNgamma IQ-ELISA Kit, TCRalpha IQ-ELISA Kit, INF-G IQ-ELISA Kit, ifng IQ-ELISA Kit, interferon gamma IQ-ELISA Kit, interferon, gamma 1-2 IQ-ELISA Kit, IFNG IQ-ELISA Kit, Ifng IQ-ELISA Kit, ifng1-2 IQ-ELISA Kit

Background: Synonyms: Interferon gamma (IFN-gamma) (Immune interferon), IFNG

Gene ID: 3458

UniProt: P01579

Pathways: Interferon-gamma Pathway, 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, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Protein targeting to Nucleus, Autophagy