At time points indicated in the text cells were washed once with PBS and resuspended in 100 μl Laemmli buffer containing 5% β-mercaptoethanol. Proteins were separated on 8% or 10% poly-acrylamide gels. Membranes were probed with antibodies recognizing IE1/2, pp28, pp65, UL44, UL84, (Virusys, 1:1000 dilution), IE2 proteins (clone 5A8.2, Millipore, 1:1000 dilution) and β-actin (SIGMA, 1:5000 dilution). All primary antibodies were detected using anti-mouse- or anti-rabbit-horseradish peroxidase (HRP) conjugated antibodies (Millipore and Cell Signaling Technology, respectively). Chemiluminescence solution (GE Healthcare) was used in each case to detect secondary antibodies using film. Relative band intensity (band intensity relative to β-actin signal in the same lane) was analyzed using ImageJ software, obtained from the National Institutes of Health (USA).
Clone 5a8
Manufactured by Merck Group
The Clone 5A8.2 is a laboratory equipment product manufactured by Merck Group. It is a specialized device designed for specific scientific applications. The core function of the Clone 5A8.2 is to perform a particular task within the laboratory environment, though the details of its intended use are not provided.
Automatically generated - may contain errors
2 protocols using clone 5a8
1
Western Blot Analysis of HCMV Proteins
2
Quantitative Immunoblotting of HCMV Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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At time points indicated in the text cells were washed once with PBS and resuspended in 100 μl Laemmli buffer containing 5% β-mercaptoethanol. Proteins were separated on 8% or 10% polyacrylamide gels. Membranes were probed with antibodies recognizing IE1/2, pp28, pp65, UL44, UL84, (Virusys, 1:1000 dilution), IE2 proteins (clone 5A8.2, Millipore, 1:1000 dilution) and β-actin (SIGMA, 1:5000 dilution). All primary antibodies were detected using anti-mouse- or anti-rabbit-horseradish peroxidase (HRP) conjugated antibodies (Millipore and Cell Signaling Technology, respectively). Chemiluminescence solution (GE Healthcare) was used in each case to detect secondary antibodies using film. Relative band intensity (band intensity relative to β-actin signal in the same lane) was analyzed using ImageJ software, obtained from the National Institutes of Health (USA).
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