Human Phospho-SMAD4 (Thr277) and Total SMAD4 PharmaGenie ELISA Kit (SBRS1973)

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SKU:
SBRS1973
Product Type:
ELISA Kit
Reactivity:
Human
Applications:
ELISA
ELISA Type:
Sandwich
Frequently bought together:

Description

Human Phospho-SMAD4 (Thr277) and Total SMAD4 PharmaGenie ELISA Kit

Product SKU:SBRS1973
Size:96T
Application:This ELISA kit recognizes Human SMAD4 phosphorylated at site Threonine-277 as well as total SMAD4.
Uniprot:Q13485
Gene ID:4089
Gene Names:DPC4 / MADH4 / SMAD4
Synonyms:Mothers against decapentaplegic homolog 4 (MAD homolog 4) (Mothers against DPP homolog 4) (Deletion target in pancreatic carcinoma 4) (SMAD family member 4) (SMAD 4) (Smad4) (hSMAD4)
Target Species:Human
Compatible Sample Types:Cell Lysates, Tissue Lysates
Design Principle:Sandwich-based
Method of Detection:Colorimetric
Quantitative/Semi-Quantitative:Semi-Quantitative
Storage/Stability:Upon receipt, the kit should be stored at -20°C. Please use within 6 months from the date of shipment.
  1. Pre-Coated 96-well Strip Microplate
  2. Wash Buffer
  3. Anti-Phospho Antibody
  4. HRP-Conjugated Secondary Antibody
  5. Assay Diluent
  6. TMB One-Step Substrate
  7. Stop Solution
  8. Lysis Buffer
  9. Positive Control Sample

Other materials and equipment required:

The Assay Genie Human Phospho-SMAD4 (Thr277) and Total SMAD4 PharmaGenie ELISA Kit (SBRS1973) 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
  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.
Product ImageJurkat cells were untreated or treated with CPT. Cell lysates were analyzed using this phosphoELISA and Western Blot.
Product ImageJurkat cells were treated with PV (Pervanadate). Cells were solubilzed at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA

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