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Cellular Western Assay Service

Cellular Western Assay Services

   —Quantitative & Cell-Based

Cellular Western Assay Services

Immunochemistry is a preferred cell-based assay technology after a biochemical screening. However, the traditional immunochemical technologies, such as conventional Western, are cumbersome and labor intensive for compound screening. To address the problem, GenScript has developed Cellular Western assay service platform based on the In-Cell WesternTM technology.

The Cellular Western is an immunocytochemical assay performed in microplate format, 96-well or 384-well plate. Target-specific primary antibodies and infrared-labeled secondary antibodies are used to detect the change of target protein level in whole cell environment. Detection of the infrared fluorescence intensity enables quantitative analysis of cellular signaling pathway. Because of fewer steps and more real cellular environment than traditional Western technology, Cellular Western is a powerful tool which brings you higher throughput and more meaningful data in compound screening.

Cellular Western Workflow

  • Direct detection of proteins in their cellular context eliminates variabilities and artifacts caused by cell lysis: More relevant results than enzyme assays with purified proteins.
  • Two simultaneous target detections: Precise quantification and accurate measurement of abundance or phosphorylation or one target by normalization against another target or DNA strain.
  • Near-infrared probes: High sensitivity for measuring small changes in protein amount or modification
  • Flexibility: Analyze multiple targets in 96- or 384-well plates, ideal for screening cell treatments or drug candidates for effects on target proteins.

Two-Color Cellular Western detection of phospho-EGFR

 

Dose response of A431 to PD168393

Fig 1.384-well two color Cellular Western with the 700 nm and 800 nm channels detects total β-actin and phosphorylated-EGFR respectively   Fig 2. Dose response of A431 cells to PD168393 as measured by specific antibody detection of phosphorylated-EGFR, using β-actin for normalization.

Two-Color Cellular Western detection of phospho-P38

Fig 3. 96-well two color Cellular Western with the 700 nm and 800 nm channels detects total ERK2 and phosphorylated-p38 (Thr180/Tyr182), respectively.

 

Dose response of Hela cells to Anisomycin

Fig 4.Dose response of Hela cells to Anisomycin as measured by specific antibody detection of phosphorylated-p38 (Thr180/Tyr182), using total ERK2 for normalization.

Related Services and Products

Quotations and Ordering:

To request a quotation, please download and complete the Cellular Western Assay Service and send it back by email or fax.

To order, please send the completed Quotation Form to us by email or fax with a formal PO (Purchase Order) or credit card information. You can also submit PO/credit card information by phone or via our Secure Online Messaging System.

Our customer service representatives are available 24 hours a day, Monday through Friday to assist you.