Interferon gamma ELISA Kit

Catalog No. ABIN6975283

Product Details Interferon gamma ELISA Kit

Target: Interferon gamma (IFNG)

Reactivity: Chicken

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 3.125 pg/mL - 800 pg/mL

Minimum Detection Limit: 3.125 pg/mL

Application: ELISA

Purpose: For the quantitative determination of chicken interferon γ (IFN-γ) concentrations in serum, plasma, tissue homogenates.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of chicken IFN-γ. No significant cross-reactivity or interference between chicken IFN-γ and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between chicken IFN-γ and all the analogues, therefore, cross reaction may still exist.

Components:

• Assay plate
• Standard
• HRP-conjugate (100 x concentrate)
• Sample Diluent
• HRP-conjugate Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Stop Solution

Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 50 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Set a Blank well without any solution.
3. Add 50 µL standard or sample to each well.
4. Add 50 µL HRP-conjugate (1x) to each well (Not to Blank well).
5. Incubate 1 hour at 37 °C
6. Aspirate and wash 5 times.
7. Add 90 μL of TMB Substrate to each well. Incubate for 20 minutes at 37 °C. Protect from light.
8. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. HRP-conjugate (1x) - Centrifuge the vial before opening. HRP-conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-conjugate + 990 μL of HRP-conjugate Diluent.
2. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
3. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 800 pg/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 150 μL of Sample Diluent into each tube (S0-S4). Use the stock solution to produce a 4-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (800 pg/mL). Sample Diluent serves as the zero standard (0 pg/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

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 Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If don't make recent use, better keep it store at HRP-conjugate -20°C. Opened kit HRP-conjugate Diluent Sample Diluent May be stored for up to 1 month at 2 - 8°C. Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for Interferon gamma ELISA Kit (ABIN6975283)

Lee, Chaudhari, Oh, Eo, Park, Jawale: "Immune responses to oral vaccination with Salmonella-delivered avian pathogenic Escherichia coli antigens and protective efficacy against colibacillosis." in: Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, Vol. 79, Issue 3, pp. 229-34, (2016) (PubMed).

Huang, Zhang, Li, Song, Yan, Xu, Li: "Eimeria maxima microneme protein 2 delivered as DNA vaccine and recombinant protein induces immunity against experimental homogenous challenge." in: Parasitology international, Vol. 64, Issue 5, pp. 408-16, (2015) (PubMed).

Huang, Zhang, Li, Song, Yan, Xu, Li: "Immune protection of microneme 7 (EmMIC7) against Eimeria maxima challenge in chickens." in: Avian pathology : journal of the W.V.P.A, Vol. 44, Issue 5, pp. 392-400, (2015) (PubMed).

Nirala, Perumal, Gohil: "Glycated serum albumin stimulates expression of endothelial cell specific molecule-1 in human umbilical vein endothelial cells: Implication in diabetes mediated endothelial dysfunction." in: Diabetes & vascular disease research, Vol. 12, Issue 4, pp. 290-7, (2015) (PubMed).

Matsuyama-Kato, Murata, Isezaki, Takasaki, Kano, Konnai, Ohashi: "Expression analysis of programmed death ligand 2 in tumors caused by the avian oncovirus Marek's disease virus." in: Archives of virology, Vol. 159, Issue 8, pp. 2123-6, (2014) (PubMed).

Zhang, Huang, Li, Sui, Wang, Liu, Xu, Yan, Song, Li: "Identification and Molecular Characterization of Microneme 5 of Eimeria acervulina." in: PLoS ONE, Vol. 9, Issue 12, pp. e115411, (2014) (PubMed).

Maroufyan, Kasim, Ebrahimi, Loh, Bejo, Zerihun, Hosseni, Goh, Farjam: "Omega-3 polyunsaturated fatty acids enrichment alters performance and immune response in infectious bursal disease challenged broilers." in: Lipids in health and disease, Vol. 11, pp. 15, (2012) (PubMed).

Moeini, Omar, Rahim, Yusoff: "Improving the potency of DNA vaccine against chicken anemia virus (CAV) by fusing VP1 protein of CAV to Marek's Disease Virus (MDV) type-1 VP22 protein." in: Virology journal, Vol. 8, pp. 119, (2011) (PubMed).

Li, Xue, Wang, Wang, Chen: "Bursopentine as a novel immunoadjuvant enhances both humoral and cell-mediated immune responses to inactivated H9N2 Avian Influenza virus in chickens." in: Clinical and vaccine immunology : CVI, Vol. 18, Issue 9, pp. 1497-502, (2011) (PubMed).

Moeini, Rahim, Omar, Shafee, Yusoff: "Lactobacillus acidophilus as a live vehicle for oral immunization against chicken anemia virus." in: Applied microbiology and biotechnology, Vol. 90, Issue 1, pp. 77-88, (2011) (PubMed).

Moeini, Omar, Rahim, Yusoff: "Development of a DNA vaccine against chicken anemia virus by using a bicistronic vector expressing VP1 and VP2 proteins of CAV." in: Comparative immunology, microbiology and infectious diseases, Vol. 34, Issue 3, pp. 227-36, (2011) (PubMed).

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:

Abbreviation: IFNG

Alias: IFG, IFI,IFN-gamma,interferon gamma

UniProt: P49708

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