Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags. |
-Run a standard curve with each assay.
-Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
-Do not cover plate with foil. |
-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity.
-Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days. |
Upstream tips |
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags. |
Protocol tips |
-Run a standard curve with each assay.
-Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells.
-Do not cover plate with foil. |
Downstream tips |
-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity.
-Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days. |
Publication protocol
To supplement EGFR IHC, we performed an ELISA on 1–3 snap-frozen rectal biopsies which were pooled. Biopsies were incubated in a uticell extraction buffer (20-30ml; Invitrogen Cat # FNN0011) supplemented with 1 mM PMSF (Invitrogen) protease inhibitor cocktail (Sigma Chemicals) for 30–45 min at 4°C and then then centrifuged at 13,000 rpm for 10 min. The supernatant underwent protein estimation with Quant-iT assay kit (Invitrogen). For EGFR quantitation, 10 μg protein lysate was assessed with Invitrogen EGFR ELISA kit (KHR9061) according to the manufacturer’s directions.
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