User:Mark X. Ling/Notebook/Micb 323/2009/03/24

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ELISA

 * dilution of TNFa standard: 118ng/mL~> 4ng/mL => 17uL stock + 484.5uL dH2O => total volume = 501.5uL
 * 1st incubation(room temperature)[1hr]@1:20->2:18
 * incubation after loading sample [1.5hr]@2:55-3:58
 * incubation after HRP [30mins]@4:02
 * only triplet was done for the supplied 8 hr LPS+IFNg
 * there were not enough sample for quadruplets
 * F12 is left empty

ELISA result:
 * additional 50uL of concentrated strep. avidin HRP added
 * 100uL initially added = too low of concentration, so we needed more.
 * Immunoplate Grid Map
 * the # indicates the sample number labeled on tube.

PAGE

 * heat all samples except the MW markers
 * incubated at 1:43 at 100oC for 2 minutes


 * gel started running at 2:16->2:56

Western Transfer

 * no comments: all procedures done according to the manual

Post lab:

 * procedure of ELISA went very smoothly
 * there was a mistake in loading the wells, but not major enough. Keep it in mind
 * didn't need to do much for Western.
 * too little reagent was used for the ELISA, no data available for our experiment
 * NOTE: result is from ELISA done by Mrs.Hinze and Jen ==> our result is not available.


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