Human Phospho-EGFR (Y1045) and Total EGFR PharmaGenie ELISA Kit (SBRS1792)

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SKU:
SBRS1792
Product Type:
ELISA Kit
Reactivity:
Human
Applications:
ELISA
ELISA Type:
Sandwich

Description

Human Phospho-EGFR (Y1045) and Total EGFR PharmaGenie ELISA Kit (SBRS1792)

Product SKU:SBRS1792
Size:96T
Application:The antibody pair provided in this kit recognizes human Phospho-EGFR (pTyr1045) + pan EGFR.
Uniprot:P00533
Gene ID:3236
Gene Names:EGFR / ERBB / ERBB1 / HER1
Pathway:HER/ErbB Signaling
Synonyms:Epidermal growth factor receptor (EC 2.7.10.1) (Proto-oncogene c-ErbB-1) (Receptor tyrosine-protein kinase erbB-1)
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. 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.
  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 Phospho-EGFR (Y1045) and Total EGFR PharmaGenie ELISA Kit (SBRS1792) 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.
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