SPINT4 Antibody - BSA Free

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• SPINT4 Antibody

  Name: Charlotte Dietzel

  Application: Immunocytochemistry

  Sample Tested: plasmid transfected HEK293T cells

  Species: Human

  Verified Customer | Posted 05/10/2023

  40x Image of SPINT4-HA 293T cells showing the non-specific binding and high background noise of NovusBio’s SPINT4 Antibody.

  Overview/Application Description: The specificity and signal/noise ratio of the anti-SPINT4 antibody was tested in immunofluorescence staining analyses of Human HEK293T cells expressing the SPINT4-HA protein. Signal seems to be non-specific (based on the degree of overlap of the SPINT4 and HA signal) with high background (based on the presence of signal in untransfected cells). Procedural Details: HEK293T cells were transfected with an expression plasmid encoding the SPINT4 protein fused to a C-terminal HA epitope (SPINT4-HA). Cells were fixed in 1.5% paraformaldehyde at room temperature, permeabilized with 0.5% Triton-X on ice, blocked using 10% fetal bovine serum in PBS and incubated with rabbit anti-SPINT4 antibodies (1:200 dilution) and with mouse anti-HA antibodies (1:200 dilution) for one hour at room temperature. Samples were washed with blocking buffer and incubated with Alexa Fluor 594-conjugated goat anti-rabbit antibodies (1:200 dilution) and with Alexa Fluor 488-conjugated goat anti-mouse antibodies (1:200 dilution) for one hour at room temperature, prior to washing in PBS and highlighting of nuclear DNA using Hoechst33342 for three minutes. Untransfected cells processed in the same way were used as controls. Coverslips were then mounted, and signal was visualized using a fluorescence microscope.

  

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