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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:

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Presentation on theme: "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:"— Presentation transcript:

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 Your sample layout (on 96 well plate): --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. Protocol for ELISA Sample Submission-singlet: sample layout 1 2 3 4 5 6 7 8 9 10 11 12 ABCDEFGHABCDEFGH

3 Example of sample submission sheet – 96 well single

4 Results will be sent to you via e-mail in the following format: Plate/Samples Name:Date: 3/10/05 x-40 87.741.818.59.04.83.12.32.1 -0.349.798.13.7 87.4 19041490.643.518.07.05.83.19.02.2 0.445.6117.73.9 43.7 8897873.628.17.5-0.417.9-1.932.51.4-6.512.8-1.8-3.2 JOE 21.9 3562183.228.07.11.47.70.90.32.5-6.511.9-0.8-2.6 10.9 1319341.39.93.80.5-0.9-0.3-2.8-2.9-7.54.1-3.6 5.5 730752.811.216.37.73.45.02.52.9-2.18.8-3.00.0 2.7 248143.918.710.210.06.16.56.34.41.751.392.97.4 JANE 1.4 811542.520.210.57.13.83.63.04.10.846.988.15.5 0.7 380 slopeintercept 2184.01-4231.6 96 well Joe, Jane x-40 y = 2184x - 4231.6 R 2 = 0.9928 -50000 0 50000 100000 150000 200000 250000 0.020.040.060.080.0100.0 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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