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Rat IFNγ Single Analyte ELISArray Kit SER45050A

Description
The Rat IFNγ Single Analyte ELISArray Kit is designed to quantitatively measure the amount of this cytokine in cell culture supernatant, serum or plasma using a conventional enzyme-linked immunosorbent assay (ELISA). Each 96-well plate (8 wells/strip, 12 strips) is coated with the protein-specific capture antibody. We screened all commercially available antibodies to identify the best capture and detection antibodies. The high sensitivity, good linearity and low background of the Single Analyte ELISArray Kits provide reliable and reproducible results for your cytokine and chemokine analyses.
Product Resources
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More information on IFNγ from NCBI:
"In contrast to interferon-α and interferon-β which can be expressed by all cells, IFN-γ is secreted by T lymphocytes and NK cells only. Also known as immune interferon, IFN-γ is the only Type II interferon. It is serologically distinct from Type I interferons and it is acid-labile, while the type I variants are acid-stable. IFN-γ has antiviral, immunoregulatory, and anti-tumour properties. It alters transcription in up to 30 genes producing a variety of physiological and cellular responses. Activation by IFN-γ is achieved by its interaction with a heterodimeric receptor consisting of IFNGR1 & IFNGR2 (interferon gamma receptors). IFN-γ binding to the receptor activates the JAK-STAT pathway. In addition, IFN-γ activates APCs and promotes Th1 differentiation by upregulating the transcription factor T-bet. IFN-γ is the hallmark cytokine of Th1 cells (Th2 cells produce IL-4). NK cells and CD8+ cytotoxic T cells also produce IFN-γ. IFN-γ suppresses osteoclast formation by rapidly degrading the RANK adaptor protein TRAF6 in the RANK-RANKL signaling pathway, which otherwise stimulates the production of NFκB."
Kit Contents / Storage Conditions/ Shelf Life
Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt.

Enough reagents are provided to process the included plate of 12 ELISA strips.

Component / Description  Quantity
BOX 1: Shipped on blue ice packs. Store at -20 ºC.
Antigen Standard (1 µg/ml)  1.5-ml tube
Detection Antibody  1.5-ml tube
Avidin-HRP Conjugate  1.5-ml tube
10% BSA  15 ml bottle
BOX 2: Shipped at ambient temperature. Store at 4 ºC.
Pre-coated Capture Antibody 8-well strips  One plate of 12 strips in a pouch
Sample Dilution Buffer Stock  60 ml bottle
Assay Buffer Stock  60 ml bottle
Wash Buffer (10 × Concentrate)  125 ml bottle
Development Solution  25 ml bottle
Stop Solution  60 ml bottle

Shelf Life: Do not use kit beyond the expiration date printed on the label.

Brief Protocol
The Brief Protocol is meant for experience users only. First-time users should refer to the User Manual.
  1. Prepare replicate serial dilutions of the Antigen Standard and your experimental samples.
  2. Pipette 50 µl of Assay Buffer into each well of the 8-well ELISA strips.
  3. Transfer 50 µl samples and/or standards to the appropriate wells of the ELISA strips.
  4. Gently shake or tap plate for 10 seconds. Incubate for 2 hours at room temperature.
  5. Washing ELISA Wells:
    Decant or aspirate well contents. Add 350 µl 1 × Washing Buffer. Gently shake or tap plate for 10 seconds. Decant or aspirate. Blot array upside down on absorbent paper to remove any residual buffer. Repeat wash twice more.
  6. Pipette 100 µl of Detection Antibody solution. Incubate 1 hour at room temperature.
  7. Wash ELISA wells as described above.
  8. Add 100 µl Avidin-HRP solution to all wells. Incubate for 30 minutes at room temperature.
  9. Wash ELISA wells for a total of 4 washes.
  10. Add 100 µl of Development Solution to each well. Incubate the plate for 15 minutes at room temperature in the dark.
  11. Add 100 µl of Stop Solution to each well. The color changes from blue to yellow.
  12. Read absorbance at 450 nm within 30 minutes of stopping the reaction. If wavelength correction is available, subtract readings at 570 nm from the reading at 450 nm.