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AdvanCell in-cell Western Kit

Total cell normalization and simultaneous detection of two biomarkers in whole cells

  • Save time – cells are directly probed, no need for time consuming lysate preparation
  • Quantitative immunofluorescence – normalize to cell number, allowing for accurate quantification and comparison of protein expression between wells
  • Detection of high and low molecular weight targets – a limitation of traditional Western blots is eliminated as high and low MW targets can easily be evaluated simultaneously
  • Detect proteins in situ in the most relevant cellular context with minimal sample processing
AdvanCell in-cell Western Kit


CAT # PRODUCT SIZE PRICE QUANTITY
K-15065-005 AdvanCell™ in-cell Western Kit, Rb700/Ms800 for 5 plates $ 415.00
(USD)
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K-15066-005 AdvanCell™ in-cell Western Kit, Rb800/Ms700 for 5 plates $ 415.00
(USD)
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L-07096-005 AdvanCell™ 96-well Tissue Culture Plates with Lids 5 each $ 175.00
(USD)
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R-03168-060 500X AdvanCell™ Total Cell Staining Solution 60μL $ 88.00
(USD)
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Description

AdvanCell™ in-cell Western Kit includes all critical reagents required to generate high sensitivity whole cell staining data for five 96-well plates. AdvanCell™ Permeabilization Solution is intended to remove membrane lipids to allow antibodies to enter the cell. This solution will also permeabilize the nuclear membrane to allow for detection of nuclear proteins. AdvanCell™ Total Cell Staining Solution is a nuclear stain, provided as a 500X solution intended to be co-incubated with primary antibodies to save time and decrease the number of assay steps. Each kit includes one Spectradye IR700 and one Spectradye IR800 conjugated secondary antibody directed against mouse or rabbit primary antibodies for simultaneous detection of two biomarkers.

 

AdvanCell in-cell Western Kit workflow

AdvanCell workflow

 

 

 

Quantitative and simultaneous measurement of two protein targets and total cell normalization

AdvanCell measurement of two protein targets

HeLa cells were co-incubated overnight with AdvanCell Total Cell Staining Solution, 1:200 Mouse anti-STAT3 (Cell Signaling Technology #9139) and 1:200 Rabbit anti-b-Actin (Millipore #MABT523). AdvanCell Total Cell Staining Solution is a nuclear stain represented by blue and imaged in the Cy3 Channel. After washing, a 1:120 dilution of goat anti-mouse SpectraDye IR700, represented by red, and a 1:120 dilution of goat anti-rabbit SpectraDye IR800, represented by green, was applied for 1 hour. After washing with 1X PBS the plate was imaged on a Sapphire Biomolecular Imager (Azure Biosystems).

Cells incubated with AdvanCell Total Cell Staining Solution and SpectraDye Secondary antibodies do not show non-specific binding of secondary antibodies to celluar proteins

AdvanCell and SpectraDye do not show non-specific binding

 

 

 

 

 

 

 

 

 

AdvanCell Total Cell Staining Solution does not interfere with primary antibody binding

AdvanCell doe not interfere with primary antibody binding

HeLa cells were co-incubated overnight with or without AdvanCell Total Cell Staining Solution, 1:200 Mouse anti-b Tubulin (Boster Bio #MA1112) and 1:200 Rabbit anti-EGFR (Cell Signaling Technology #4267). After washing, a 1:120 dilution of goat anti-mouse SpectraDye IR700, represented by red, and a 1:120 dilution of goat anti-rabbit SpectraDye IR800, represented by green, was applied for 1 hour. After washing with 1X PBS the plate was imaged on a Sapphire Biomolecular Imager (Azure Biosystems).

AdvanCell Total Cell Staining Solution stains cells linearly from 0-25,000 cells

AdvanCell stains cells linearly from 0-25,000 cells



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