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Protocol for ELISA Sample Submission-96 well : General Procedures

Protocol for ELISA Sample Submission-96 well : General Procedures Primary assays: x-40 & x-42 . (1-40, 1-42, & 1-total on special request). Linear range: 1pM (4pg/ml) - 110pM (500pg/ml). 110uL /well is needed for each assay. If both x-40 and x-42, then submit 210 uL.

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Protocol for ELISA Sample Submission-96 well : General Procedures

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  1. Protocol for ELISA Sample Submission-96 well: General Procedures • Primary assays: x-40 & x-42. (1-40, 1-42, & 1-total on special request). • Linear range: 1pM (4pg/ml) - 110pM (500pg/ml). • 110uL/well is needed for each assay. If both x-40 and x-42, then submit 210 uL. • 2G3 and 21F12 capture antibodies used for x-40 and x-42 assays, respectively; 266B antibody used for detection. • Submit samples on 96 well plate!! • Please DO NOT use tissue culture plates. • Please leave 2 rows empty for peptide standard. • Always submit a submission sheet ahead of time. Even if you talk to Jackie about preparing a submission, you should submit a sheet at that time. This is the ONLY way that Jackie can plan ahead and prepare for assays. Submission sheets can be emailed to Jackie or dropped off at Jackie’s desk.

  2. Protocol for ELISA Sample Submission-singlet: sample layout • --For 1 assay submit 110ul/sample; for 2 assays(x-40 & x-42) submit 210ul/sample • --Add samples to a 96 well plate, starting at C1 and moving across, as shown on left below. • --Fill all 12 wells in each row before starting another row. • --Fill up to 6 rows. Your sample layout (on 96 well plate): 1 2 3 4 5 6 7 8 9 10 11 12 A B C D E F G H

  3. Example of sample submission sheet – 96 well single

  4. Plate/Samples Name: Date: x-40 96 well Joe, Jane 3/10/05 x-40 -0.3 49.7 98.1 3.7 87.7 41.8 18.5 9.0 4.8 3.1 2.3 2.1 250000 x-40 87.4 0.4 45.6 117.7 3.9 190414 90.6 43.5 18.0 7.0 5.8 3.1 9.0 2.2 43.7 JOE 88978 73.6 28.1 7.5 -0.4 17.9 -1.9 32.5 1.4 -6.5 12.8 -1.8 -3.2 200000 21.9 35621 83.2 28.0 7.1 1.4 7.7 0.9 0.3 2.5 -6.5 11.9 -0.8 -2.6 150000 10.9 13193 41.3 9.9 3.8 0.5 -0.9 -0.3 -2.8 -2.9 -7.5 4.1 -3.6 -3.6 5.5 7307 52.8 11.2 16.3 7.7 3.4 5.0 2.5 2.9 -2.1 8.8 -3.0 0.0 100000 2.7 JANE 2481 43.9 18.7 10.2 10.0 6.1 6.5 6.3 4.4 1.7 51.3 92.9 7.4 y = 2184x - 4231.6 1.4 50000 8115 42.5 20.2 10.5 7.1 3.8 3.6 3.0 4.1 0.8 46.9 88.1 5.5 2 R = 0.9928 0.7 380 0 slope intercept 0.0 20.0 40.0 60.0 80.0 100.0 2184.01 -4231.6 -50000 Results will be sent to you via e-mail in the following format: Standards-top two rows in yellow Blank controls-highlighted in gray Conditioned media controls and plasma controls-between blank controls Your samples-will all be highlighted in one color with your name next to the samples

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