Equipment:
Affinity chromatography column (Bio-Rad 731-1550 Poly-Prep column, 0.8×4 cm)
Fraction collector (approximately 25 additional clean tubes are needed)
Reagents:
Metal Chelate Affinity Chromatography Gel (Ni-NTA agarose, QIAGEN 30210), 1 mL
#The nitrilotriacetic acid (NTA) groups are covalently attached to the cross-linked agarose matrix, allowing nickel ion chelation. The gel is pre-saturated with nickel ions before use.
Buffer B-300/0: 50 mM Na₃PO₄, pH 8.0; 0.3 M NaCl
#Before use, add 10 mM β-mercaptoethanol to the buffer.
Buffer B-300/20: Buffer B-300/0 with 20 mM imidazole added
Buffer B-300/250: Buffer B-300/0 with 250 mM imidazole added
Procedure:
1. The Ni-NTA agarose gel (1 mL) has been packed into the column, forming a light blue gel bed. Place the column in a vertical position on the column holder. Remove the packing plug at the bottom and rinse the column with 20 mL of Buffer B-300/0. Close the outlet to prepare for sample loading.
2. Carefully remove the buffer above the gel surface. Take the sample (IEX) and bring it to room temperature. Slowly apply the sample onto the top of the gel using a pipette, ensuring not to disturb the gel surface. Allow the sample to flow through the column, collecting the eluate in 2.5 mL fractions per tube.
3. Once all the sample has passed through the column, close the outlet. Then slowly add Buffer B-300/20 to the column and open the outlet to collect the effluent. Perform this wash with 30 mL of Buffer B-300/20.
4. Next, wash the column with 30 mL of Buffer B-300/250 using the same procedure as step 3. Collect each fraction accordingly.
5. After collection, measure the protein concentration and enzyme activity of each fraction. Select the fractions with the highest enzyme activity. Combine these fractions to obtain the total volume (AF). Reserve 100 μL of the final preparation for further analysis or storage.
(Word count: 520)LED Front Lit Halo Lit Letters
We are a factory direct outlet in A.D industry, more than 13 years experience.
Main produce advertising signs, such as lightbox, luminous letters, directory and Wayfinding Sign, aluminum profile sign,waste bin,mail/parcel drop box and some project sign.
Make it easily to our customer
Contact us →A reference picture or your design→Size information→Confirm the size and material→You will get our best offer.
FAQ
Q: Are you a trading company or factory?
A: We are factory.
Q: What is the MOQ?
A: No MOQ required normally, that means you can always ask for a sample first. But remember the sample freight is at your own cost.
Q: Do I have to have a design to inquire?
A: Yes! A well made design/artwork is the most helpful thing to get an exact quote. However, if you really do not have a design...just let us know your idea, we'll cover it.
Product name: LED Frontlit Halo Lit Letters
Marterial: stainless steel+acrylic
Size: customize
Color: customize
Warranty: 1 year
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Chengdu GodShape Sign Co., Ltd , https://www.signsgs.com