Difference between revisions of "Part:BBa K2876014"

Revision as of 22:04, 17 October 2018

IL1B

This sequence produces protein interleukin-1B, a human protein produced during fevers. For our project we detected IL1B with two single-chain antibodies fused to transcription initiating proteins (BBa_K2876001, BBa_K2876002). We validated our IL1B with an ELISA (https://www.thermofisher.com/elisa/product/IL-1-beta-Human-ELISA-Kit/BMS224-2) which proved production of IL1B, and that the IL1B produced was folded properly so that antibodies could bind. Validation of antibody binding was essential to ensure that IL1B could function as the target protein in our Single-Chain Antibody Prokaryotic Two Hybrid Detection System (Figure 4).

Figure 1: A graph of the absorbance at 540 nm from an ELISA of DH5-alpha cells transformed to produce IL1B (in orange) compared to the IL1B-Standard (in blue) and the negative control of untransformed DH5-alpha cells (in grey). Protein was extracted using a B-PER protocol.

Figure 2: A photo of the IL1B ELISA showing color change as a function of IL1B presence. Columns 1,2, and 6 show the ELISA standards, from least (row A) to most (row H) diluted. Columns 3, 4, and 5 rows A-D are DH5-alpha cells transformed with our IL1B plasmid. Columns 3, 4, and 5 row E are untransformed DH5-alpha cells.

Figure 3: A photo of the IL1B ELISA showing color change as a function of IL1B presence, after stop-solution added. Columns 1,2, and 6 show the ELISA standards, from least (row A) to most (row H) diluted. Columns 3, 4, and 5 rows A-D are DH5-alpha cells transformed with our IL1B plasmid. Columns 3, 4, and 5 row E are untransformed DH5-alpha cells.

Sequence and Features