Human/Mouse Phospho-Stat 1 (S727) and Total Stat 1 PharmaGenie ELISA Kit (SBRS1981)

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Human/Mouse Phospho-Stat 1 (S727) and Total Stat 1 PharmaGenie ELISA - Kit Data

Product SKU:SBRS1981
Application:The antibody pair provided in this kit recognizes human and mouse Phospho-Stat1 (pSer727) + pan Stat1.
Gene ID:6772
Gene Names:STAT1
Pathway:JAK/STAT Signaling
Synonyms:Signal transducer and activator of transcription 1-alpha/beta (Transcription factor ISGF-3 components p91/p84)
Target Species:Human, Mouse
Compatible Sample Types:Cell Lysates, Tissue Lysates
Design Principle:Sandwich-based
Method of Detection:Colorimetric
Storage/Stability:Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Kit Components:

1.Pre-Coated 96-well Strip Microplate
2.Wash Buffer
3.Anti-Phospho Antibody
4.Anti-Pan Antibody
5.HRP-Conjugated Secondary Antibody
6.Streptavidin-Conjugated HRP
7.Assay Diluent
8.TMB One-Step Substrate
9.Stop Solution
10.Lysis Buffer
11.Positive Control Sample

Other materials and equipment required:

The Assay Genie Human/Mouse Phospho-Stat 1 (S727) and Total Stat 1 ELISA Kit will require other equipment and materials to carry out the assay. Please see list below for further details.

  • Distilled or deionized water
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare sample dilutions
  • Protease and Phosphatase inhibitors
  • Precision pipettes to deliver 2 ul to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • Benchtop rocker or shaker
  • Microplate reader capable of measuring absorbance at 450 nm

Kit Protocol:

1.Prepare all reagents and samples as instructed in the manual.
2.Add 100 ul of sample or positive control to each well.
3.Incubate 2.5 h at RT or O/N at 4 °C.
4.Add 100 ul of prepared primary antibody to each well.
5.Incubate 1 h at RT.
6.Add 100 ul of prepared 1X HRP-Streptavidin to each well.
7.Incubate 1 h at RT.
8.Add 100 ul of TMB One-Step Substrate Reagent to each well.
9.Incubate 30 min at RT.
10.Add 50 ul of Stop Solution to each well.
11.Read at 450 nm immediately.