Purification and Quantification of TGFβ1

Samples were prepared as follows: 10 ml 100 ng/ml active TGFβ1 (Bio-Techne) in PBS. 10 ml 50 mg/ml VA extract in PBS. 5 ml 100 ng/ml active TGFβ1 in PBS plus 5 ml 50 mg/ml VA extract in PBS, mixed for 4 h on a rotator at RT. Samples were loaded using a 50 ml Superloop (Cytiva) and run on an ACTApurifier 10 system (Cytiva) using the equipment, reagents and programme described^{29 (link)}. Fractions were assayed for TGFβ1 using an ELISA as described.

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Lynham S., Freile F.G., Puri N.M., O’Reilly N., Mitchell G.H., Wells T.N., Willcox M, & Beatson R. (2021). Identification of chlorophyll a-b binding protein AB96 as a novel TGFβ1 neutralizing agent. Scientific Reports, 11, 7740.

Publication 2021

active TGFβ1

Elisa Tgfβ1

Corresponding Organization :

Other organizations : King's College London, University of Essex, Medicines for Malaria Venture, University of Southampton

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Variable analysis

independent variables

10 ml 100 ng/ml active TGFβ1 (Bio-Techne) in PBS
10 ml 50 mg/ml VA extract in PBS
5 ml 100 ng/ml active TGFβ1 in PBS plus 5 ml 50 mg/ml VA extract in PBS, mixed for 4 h on a rotator at RT

dependent variables

Fractions were assayed for TGFβ1 using an ELISA

control variables

PBS as the solvent for the active TGFβ1 and VA extract solutions
Rotator at room temperature (RT) for the mixed sample

positive controls

None specified

negative controls

None specified

Annotations

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