Coding

Part:BBa_K2876014

Designed by: Eleanor Glockner   Group: iGEM18_Stanford   (2018-10-10)
Revision as of 22:02, 17 October 2018 by Ijustice (Talk | contribs)


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.

IL1B graph.png

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.

IL1B ELISA Pre-stop.jpg


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.

IL1B ELISApoststop.png


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 603
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//collections/immune_regulation/inflammatory
Parameters
None