Preparative Membrane Protein Folding Column Set (column #1)

Mfr.No.	MFC01
Description	The Preparative Membrane Protein Folding Column Sets are used for preparative membrane protein folding after the folding condition has been identified by the Spin-column Membrane Protein Folding Screen Kit (Catalog # MFC01-20). The column number represents the specific folding condition. Each Column Set includes 4 identical preparative protein folding columns and reagents for folding about 5 mg of urea -solubilized inclusion body proteins. The Preparative Membrane Protein Folding Column Sets includes 5 ml of Reagent A or Reagent B, 8.4 ml of Reagent C, 15.4 ml of Solution S and 4 membrane protein folding columns. Reagent A is for column #1 to #10; Reagent B is for column #11-20.
Protein science	Membrane protein folding

Reference	1. Pelletier DM. Hemoglobin binding protein from Actinobacillus pleuropneumoniae: a novel method for extraction and isolation. McGill University, Montreal Q.C. Graduate thesis, (2007). 2. Giacani L., et al, Transcription of TP0126, Treponema pallidum Putative OmpW Homolog, Is Regulated by the Length of a Homopolymeric Guanosine Repeat, Infection and Immunity 83: 2275- 2289 (2015).

Product Manual	Datasheet
Protocols Description	PROTEIN FOLDING PROCEDURE (1) Inclusion body solubilization: Solubilize the inclusion bodies in 20 mM Tris-HCl, pH 7.0, 8 M urea, 10 mM DTT, 2 mM EDTA by stirring at room temperature for 4 hr. Centrifuge the solubilization material at 125,000 x g for 30 min to remove any insoluble materials. Adjust the protein concentration to about 2 – 4 mg /ml. (2) Sample preparation: To make the loading sample for each column, mix 1.25 ml of the solubilized inclusion bodies with 1.25 ml of Reagent A or B (Reagent A for column #1 -10, Reagent B for column #11-20). Incubate the loading sample at room temperature for 2 hr. (3) Protein folding: Step 1: Cut off the bottom tip of each column and let the buffer run through the column, load 2.5 ml of the loading sample. Let the sample run completely into the column. Provide a little pressure on the top of the column with the column cap if the flow stops. Discard the flow-through. Step 2: Elute the protein with 3.5 ml of Solution S. The Solution S number matches the column number. Collect and incubate the eluent at 4ºC for 2 hr. Discard the columns. Step 3: Mix 2 ml of Reagent C with the 3.5 ml of eluent from per column and incubate the solution at 4ºC for 2 hr to overnight. Remove the precipitate (if there is any) by centrifugation. PROTEIN PURIFICATION AFTER FOLDING The folded protein can be purified by affinity, ion-exchange or gel filtration column chromatography. Some proteins are sensitive to low salt. Concentration by ultrafiltration may also cause concentration of the detergent that may denature the folded membrane protein. To be cautious, brief dialysis of the protein solution against a buffer with a moderate salt concentration at 4ºC is recommended. Following is a Q-Sepharose column purification protocol: (1) Dialyze the protein solution against 40 volumes of 20 mM Tris-HCl, pH 8.5, 50 mM NaCl, 1 mM dodecyl maltoside, 2 mM EDTA buffer at 4ºC for 2 to 4 hr. Remove any precipitates by centrifugation. (2) Equilibrate a Q-Sepharose column with a low salt buffer (the same as the dialysis buffer). Load the dialyzed protein solution. Wash the loaded column with 10 column volumes of the same buffer. Elute the protein with a salt gradient from 50 mM to 1 M. (3) Any further protein purification step may follow as purification of native membrane proteins.

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Combination SKU	Supplier No.	Specifications	Your price
BHE16300325-1	MFC01	column #1	$366.64	Add to Cart
BHE16300325-2	MFC10	column #10	$366.64	Add to Cart
BHE16300325-3	MFC11	column #11	$366.64	Add to Cart
BHE16300325-4	MFC12	column #12	$366.64	Add to Cart
BHE16300325-5	MFC13	column #13	$366.64	Add to Cart
BHE16300325-6	MFC14	column #14	$366.64	Add to Cart
BHE16300325-7	MFC15	column #15	$366.64	Add to Cart
BHE16300325-8	MFC16	column #16	$366.64	Add to Cart
BHE16300325-9	MFC17	column #17	$366.64	Add to Cart
BHE16300325-10	MFC18	column #18	$366.64	Add to Cart
BHE16300325-11	MFC19	column #19	$366.64	Add to Cart
BHE16300325-12	MFC02	column #2	$366.64	Add to Cart
BHE16300325-13	MFC20	Column #20	$366.64	Add to Cart
BHE16300325-14	MFC03	column #3	$366.64	Add to Cart
BHE16300325-15	MFC04	column #4	$366.64	Add to Cart
BHE16300325-16	MFC05	column #5	$366.64	Add to Cart
BHE16300325-17	MFC06	column #6	$366.64	Add to Cart
BHE16300325-18	MFC07	column #7	$366.64	Add to Cart
BHE16300325-19	MFC08	column #8	$366.64	Add to Cart
BHE16300325-20	MFC09	column #9	$366.64	Add to Cart

Preparative Membrane Protein Folding Column Set (column #1)

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