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IFN-gamma Signaling Pathway ELISA Sampler Pack

SKU:
SKFI0057
Product Type:
ELISA Kit
ELISA Type:
ELISA Sampler Pack
Size:
4 X 48 Assays
Reactivity:
Human
Analytes:
JAK1
IFNGR1
STAT1
JAK2
  • Human JAK1 ELISA Kit
  • Human STAT1 ELISA Kit
  • Human JAK2 ELISA Kit
  • Human IFNGR1 / IFN gamma receptor 1 ELISA Kit
€1,100
Frequently bought together:

Description

IFN-γ Signaling Pathway ELISA Sampler Kit

The IFN Signaling Pathway ELISA Kit Pack is a comprehensive and reliable tool for the quantitative measurement of various proteins involved in the interferon signaling pathway. This kit is specifically designed for the accurate detection of key proteins associated with interferon signaling in cell lysates, tissue homogenates, and other biological samples.The interferon signaling pathway plays a critical role in the body's immune response to viral infections, autoimmune diseases, and cancer. By accurately measuring the levels of key proteins in this pathway, researchers can gain valuable insights into the mechanisms underlying these diseases and develop targeted therapies for intervention.

With high sensitivity and specificity, this ELISA kit pack provides researchers with the tools they need to confidently study the interferon signaling pathway in a wide range of research applications. Whether investigating the molecular mechanisms of disease or evaluating the efficacy of potential therapeutics, this kit pack offers reliable and reproducible results to support your research goals.

IFN-γ Signaling Pathway ELISA Kit Pack

IFN-γ Signaling Pathway ELISA Kit Pack from Assay Genie allows researchers to begin their investigation into IFN-γ Signaling Pathway via analysing JAK1, IFNGR1 / IFN gamma receptor 1, STAT1, JAK2 in a 48 assay format, thus allowing researchers to identify and validate analytes within the pathway.

Pack ContentsSizeReactivitySensitivityRange ELISA Type
Human JAK1 ELISA Kit48 AssaysHuman0.469ng/ml0.781-50ng/mlSandwich ELISA
Human IFNGR1 / IFN gamma receptor 1 ELISA Kit48 AssaysHuman0.094ng/ml0.156-10ng/mlSandwich ELISA
Human STAT1 ELISA Kit48 AssaysHuman0.094ng/ml0.156-10ng/mlSandwich ELISA
Human JAK2 ELISA Kit48 AssaysHuman0.094ng/ml0.156-10ng/mlSandwich ELISA

Each individual ELISA kit in this pack will come with the following components:

Component Quantity Storage
ELISA Microplate (Dismountable) 4-12 strips 4°C for 6 months
Lyophilized Standard 1 4°C/-20°C
Sample/Standard Dilution Buffer 10ml 4°C
Biotin-labeled Antibody(Concentrated) 60ul 4°C (Protect from light)
Antibody Dilution Buffer 5ml 4°C
HRP-Streptavidin Conjugate(SABC) 60ul 4°C (Protect from light)
SABC Dilution Buffer 5ml 4°C
TMB Substrate 10ml 4°C (Protect from light)
Stop Solution 5ml 4°C
Wash Buffer(25X) 15ml 4°C
Plate Sealer 3 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 - g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

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