Western Blot check of SMC1fs production in cell lysate with varied conditions

2015-01-13

azim58 - Western Blot check of SMC1fs production in cell lysate with
varied conditions

The purpose of this experiment was to make sure that bacteria produced
the expected protein (SMC1fs) when the bacteria were induced in a 96 well
plate format. The bacteria were subjected to varying conditions to
determine what the best conditions were.

Expected GST-SMC1fs size: 29.2 kD
Dye Used: AF647
Excitation wavelength: 633 nm
Emission wavelength: 670 nm
Ladder used: Precision Plus Dual Color Standard

Samples all at 30 C in this image

"C:\Users\kurtw_000\Documents\kurt\storage\CIM Research Folder\DR\2014\01-24-2014d0911\01-24-2014d0911\memb1_smc1_fs_cell_lysate_clustering_10-10-11.PNG"

Note: I think this membrane with the 6hr samples does not look as good
because the sponge used to dry it fell onto the counter beforehand. The
bands are still visible and quite strong though.

Primary Sera Source: Purified Rabbit h-SMC1fs 0.73 mg/mL 8/7/07 (diluted
1:1,000)
Secondary Sera Source: Goat anti rabbit IgG h+l 1 mg/mL (diluted 1:10,000)
Tertiary Source: AF647 2 mg/mL (diluted 1:10,000)

Mouse sera was also tested
These mice were only genetically immunized

Note: The TEV-thioredoxin-His tag appears to bind the antibody quite well
too, or there was too much lysozyme in the negative control.

Supporting Files -&gt;

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