Text for indexing 8-16-12 dot 1146
2015-01-13azim58 - Text for indexing 8-16-12 dot 1146
CSGNRVWWYTKQ. . peptide removed
Images of Membranes
Tumor Naive No Antibody
Minimum Displayed Value: 200
Maximum Displayed Value: 6000
Note: Both the naive and tumor samples had about the same intensity for
some of the 6 random peptides. Both types of sera
displayed high intensities for the NRVWW, YNPIW, and VNGEY peptides. The
VNGEY peptide was the only peptide for which
the tumor sera had higher intensity than the naive sera. Both sera types
had a low intensity for the negative control SIINFEKL
peptide. One thing to keep in mind is that the tumor sera was essentially
diluted 1 million fold (1000 fold dilution during the
antibody purification since the peptide was eluted in a 1 mL fraction and
1000 fold for the dot blot assay). The naive sera was only
diluted 1000 fold for the dot blot assay. Therefore, the two sera types
are not directly comparable. If I want a better comparison,
I could redo the dot blot assay by diluting the naive sera 1 million fold
or even purify the naive sera using the same random peptide
tentagel bead method, and then perform the dot blot.
Dot Blot Summary 8-16-12
Tumor sera was purified using a mixture of Tentagel beads with 6 random
peptides. The 1st 1 mL elution fraction was then diluted
1 to 1000 for a dot blot. Naive sera was used directly by diluting 1 to
1000 for a dot blot without the antibody purification. A membrane
with no applied antibody (no primary or secondary) was also scanned on
the Typhoon. Any AF647 direct labelled secondary antibodies
were detected, and the results are summarized below.