Interferon gamma ELISA Kit

Catalog No. ABIN7014028

Product Details Interferon gamma ELISA Kit

Target: Interferon gamma (IFNG)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 2.34 pg/mL - 150 pg/mL

Minimum Detection Limit: 2.34 pg/mL

Application: ELISA

Purpose: For quantitative detection of IFN-γ in serum, plasma, tissue homogenates and other biological fluids.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of IFN-γ. No significant cross-reactivity or interference between IFN-γ and analogues were observed.

Sensitivity: 1.41 pg/mL

Grade: High Sensitivity

Components:

• Pre-coated, ready to use 96-well strip plate
• Plate sealer for 96 wells
• Standard
• Sample/Standard Dilution Buffer
• Assay Diluent
• Biotin-labeled Antibody (Concentrated)
• HRP-Streptavidin (HRP-SA)
• Biotin System (BS)
• BS Dilution Buffer
• TMB Substrate
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 50 μL

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 50μL Assay Diluent to each well
3. Add 50μL standard or sample to each well. Incubate 90 minutes at 37 °C,
4. Aspirate and wash 2 times,
5. Add 100μL Biotin-labeled antibody to each well. Incubate 1 hour at 37 °C,
6. Aspirate and wash 2 times,
7. Add 100μL BS Working Solution to each well. Incubate 15 minutes at RT,
8. Aspirate and wash 3 times,
9. Add 100μL HRP-SA to each well. Incubate 30 minutes at 37 °C,
10. Aspirate and wash 3 times,
11. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
12. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

Bring all reagents and samples to room temperature for 20 minutes before use.

1. Wash Buffer: Dilute 30 mL Concentrated Wash Buffer to 750 mL Wash Buffer with deionized or distilled water. Put unused solution back at 2-8 °C.
   Note: If crystals have formed in the concentrate, you can warm it with 40 °C water bath (Heating temperature should not exceed 50 °C) and mix it gently until the crystals have completely been dissolved. The solution should be cooled to room temperature before use.
2. Standards:
   a) Add 1 mL Sample Dilution Buffer into one Standard tube (labeled as zero tube), keep the tube at room temperature for 10 minutes and mix them thoroughly.
   Note: If the standard tube concentration higher than the range of the kit, please dilute it and label as zero tube.
   b) Label 7 EP tubes with 1/2, 1/4, 1/8, 1/16, 1/32, 1/64 and blank respectively. Add 0.3 mL of the Sample Dilution Buffer into each tube. Add 0.3 mL of the above Standard solution (from zero tube) into 1st tube and mix them thoroughly. Transfer 0.3 mL from 1st tube to 2nd tube and mix them thoroughly, and so on. Sample Dilution Buffer is used for the blank control.
   Note: It is best to use Standard Solutions within 2 hours.
3. BS Working Solution: Prepare it within 15 minutes before experiment.
   a) Calculate required total volume of the working solution: 0.1 mL/well x quantity of wells. (Allow 0.1-0.2 mL more than the total volume.)
   b)Dilute the BS with BS Dilution Buffer at 1:100 and mix them thoroughly. (i.e. Add 1 μL of BS into 99 μL of BS Dilution Buffer.)
   Note: If crystals have formed in the BS, you can warm it with water (temperature should not exceed 30 °C) and mix it gently until the crystals have completely been dissolved.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-Assay: CV<8% Inter-Assay: CV<10%

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

Expiry Date: 6 months

Target Details for Interferon gamma

Target: Interferon gamma (IFNG)

Alternative Name: Interferon gamma (IFNG Products)

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

Background: Interferon gamma (IFN-γ) is a cytokine critical to both innate and adaptive immunity, and functions as the primary activator of macrophages, in addition to stimulating natural killer cells and neutrophils. Interferon- gamma (IFN-gamma), also known as type II or immune interferon exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine. Mature human IFN-gamma exists as a non-covalently linked homodimer of 20-25 kDa variably glycosylated subunits. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interacts with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction. IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activat ion of monocytes and macrophages, up-regulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN -gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis.

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