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Human/Mouse/Rat Phospho-Stat 3 (Y705) ELISA

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SKU:
RHHU0001
$738

Description

 

Accession Number:P40763
Gene ID: 6774
Gene Symbol: APRF, STAT3
Protein Name & Synonyms: Signal transducer and activator of transcription 3 (Acute-phase response factor)
Pathway: JAK/STAT Signaling
Specificity: The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Stat3 (pTyr705).
Number of Targets Detected: 1
Species Detected: Human, Mouse, Rat
Compatible Sample Types: Cell & Tissue Lysates
Design Principle: Sandwich based ELISA
Size: 96 assays
Kit Components: Pre-Coated 96-well Strip Microplate
  Wash Buffer
  Anti-Phospho Antibody
  HRP-Conjugated Secondary Antibody
  Assay Diluent
  TMB One-Step Substrate
  Stop Solution
  Lysis Buffer
  Positive Control Sample
Other Materials Required: 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 µl 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
Protocol Outline: 1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
  7. Incubate 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.
Storage: 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.
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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