independent validation review: mcherry tag antibody (stj34373)
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Improving Quality And Reproducibility Through Antibody Validation
NEW INDEPENDENT VALIDATION RESULTS
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Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: mCherry Tag antibody (STJ34373)
Antibody: STJ34373. mCherry Tag AntibodyMethod for validation: Immunofluorescence Positive Control: A549 cells transfected with mCherry vectorNegative Control: untransfected A549 cellsResult of validation: Specific fluorescent signal was detected in the positive control, and no signal was detected in either the negative controls or the experimental controls
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Figure: A549 cells transfected with mCherry vector stained with mCherry antibody (green). mCherry itself also gives fluorescence signal (red). Bottom-right panel shows merged image from blue, green, and red channels.
Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: mCherry Tag antibody (STJ34373)
Primary antibody: Product Catalog No: STJ34373. mCherry Tag AntibodyProvider: St John's LaboratoryDilution ratio: followed provider's recommendationSecondary antibody: Antibody: Goat anti-Mouse IgG+IgM Antibody (DyLight 488)Controls:Positive control: A549 cells transfected with mCherry vector pLV-mCherry stained with mCherry antibodyNegative control: Untransfected A549 cells stained with mCherry antibodySecondary Only control: A549 cells transfected with mCherry vector and stained with Goat anti-mouse IgG+IgM Antibody (DyLight 488) only.Isotype control: A549 cells transfected with mCherry vector and stained with isotype control mouse IgG1 (Cusabio).
www.stjohnslabs.com
Improving Quality And Reproducibility Through Antibody Validation
ANTIBODY VALIDATION REPORT: mCherry Tag antibody (STJ34373)
Plating the cells:200k of A549 cells were plated in each well in a 24-well plate and cultured overnight in serum-free media in an incubator at 37ºC with 5% CO2.Transfection: Cells were transfected with pLV-mCherry using Lipofectamine 2000 according to manufacturer's protocol and grown overnight before fixing.Fixing/permeablization/staining:Cells were fixed with 4% PFA for 15 min at RT.Cells were then permeablized with 0.1% Triton-X100 for 30 min.Afterwards, cells were blocked using 1% BSA for 30 min.Cells were incubated with mCherry antibody (1:200, 1mg/ml) or isotype control (1:400, 2mg/ml) overnight at 4ºC.Cells were washed with PBS 3X at 5 min each.Cells were then incubated with Goat anti-mouse IgG+IgM Antibody (DyLight™ 488) (1:200) for 30 min and then counter stained with Hoechst 33342.Fluorescence signals were observed using a Leica fluorescence microscope.
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Improving Quality And Reproducibility Through Antibody Validation
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