SDS-PAGE / Western Blotting Lab

Note: Use this quantitative data to think about the results of this experiment. You do not need to include the photos given here in your lab report... just use the ones that you generated yourselves.

	(click image for larger view)	(click image for larger view)
	Coomassie Blue Stained Gel The left-most lane is the MW markers; see below for sample order.	Western Blot  (Probed using an anti-BSA antibody)

Note:

The blot should be the mirror image of the gel (think about the process). However, I used a image editor to "flip" the blot over so that the lanes appear in the same order as they do on the gel.

I also used the image analysis software that comes with the gel imager to quantitate the intensity of staining in both photos. This data is useful in thinking about your results. Specifically, think about the meaning of the ratio of the intensities.

The results are as follows:

			Net Intensity
	Lane	ID	Gel	Blot	Ratio
	0	MWM*	--	--	--
	1	cow	22566	27842	1.2
	2	horse	33336	13621	0.4
	3	sheep	25338	11550	0.5
	4	donkey	27737	17173	0.6
	5	chicken	27157	4730	0.2

*MWM = Molecular Weight Markers (gel only)

phosphorylase B ­ 97 kDa
bovine serum albumin - 66 kDa
ovalbumin - 45 kDa
carbonic anhydrase -29 kDa