Simple and rapid extraction of IgG

The application of ammonium sulfate salting-out and DEAE-cellulose chromatography to purify purified IgG not only makes the operation complicated and takes a long time, but also greatly increases the sample volume during the treatment process, and the denaturation is serious during dialysis, which easily causes the antibody titer to decrease. In order to overcome this shortcoming, especially when a large amount of IgG is extracted, the following simple method is often adopted.

1. DEAE-cellulose direct extraction method

Take the sample directly through the DEAE-cellulose column. The simplest beaker method is described below.

â‘´ Put a certain amount of DEAE52 into the beaker, add 0.01Mol / L pH8.0PB buffer solution, let stand for 30min, remove the supernatant fine particles, and repeat again.

⑵ Filter with Büchner funnel (with two layers of filter paper inside).

(3) Add the ingredients in the ratio of 5g wet weight DEAE-cellulose plus 1ml serum and 3ml distilled water mixture, and stir well.

⑷Place at 4 ℃ for 1h, stirring several times in the middle.

⑸ Suction filtration with Büchner funnel, then rinse cellulose with 0.01Mol / L pH8.0PB buffer solution, and suction filtration. The filtrate is the extracted IgG liquid.

2. DEAE-Sephadex A-50 is a weakly basic anion exchanger. After the Cl-type is converted to OH-type by NaOH, it can adsorb acidic proteins. The rest of the serum are acidic proteins except γ-G which is a neutral protein. When the pH of the solution is 6.5, acidic proteins are adsorbed by DEAE-Sephadex A-50, and only γ-G remains in the solution. Therefore, γ-G can be extracted using this principle. This extraction process is greatly shortened, the sample volume before and after purification does not change much, and the resulting γ-G is not denatured. After four treatments, its purity is also good. The disadvantage is that the collection volume is small and the loss is large.

â‘´ DEAE-SephadexA-50 treatment. Take a few grams of DEAE-SephadexA-50, suspend in distilled water, and after 1h, pour off the upper small particles, then treat with 0.50Mol / L NaOH for 1h, wash with distilled water until neutral, then treat with 0.5 Mol / L HCl for 0.5h, wash To neutrality, finally balance with 0.01Mol / L pH6.5PB liquid and drain to dry.

⑵ Take a certain amount of serum, add an equal amount of 0.01Mol / L pH6.5PB liquid, then add 1/4 of the liquid volume of filtered DEAE-SephadexA-50, mix, and place at 4 ℃ for 1h, stirring, suction filtration, Collect the filtrate.

(3) Treat the filtrate with DEAE-Sephadex A-50 of the same weight. Repeat three times. (Four times in total). The filtrate obtained is the γ-G product.

⑷ Recovery of DEAE-SephadexA-50. It is eluted with 0.5Mol / L Na2HPO4, filtered with suction until there is no protein in the filtrate (OD280 ﹤ 0.04), and then washed with distilled water until neutral to be recovered for use.

Gas Fire Pit

Black gas firepit is a versatile gas-burning firepit, perfect for any yard or patio and indoor. It`s easy to move from one spot to another, and it is big enough for friends party and also great for just a small family affair. This gas firepit is equipped with four pieces decorative glasses.Gas regulator from different countries can be supplied. and they all have certification for different countries. The decoration covering the flame can be high temperature resistant glass balls and volcanic stones.It is the most common heating tool in modern families. Compared with the liquid gas heaters, electric heaters and coal-fired heaters used in traditional families, it is more convenient and fast, and the safety is relatively high.In the cold winter, the temperature drops sharply, more than ten degrees below zero is very common, so a variety of heating equipment has become an essential product for people. Gas heater is a very popular heating equipment, a lot of people have in the home to buy and use. However, consumers who need to buy heating equipment often do not have a special understanding of these products. Today, we will introduce some product features and advantages of gas heaters to help more consumers buy the most suitable products when they buy gas heaters.

Gas Fire Pit,Propane Fire Pit,Outdoor Gas Fire Pit,Gas Fire Pit Table

Henan Jinbailai Industrial Co.,Ltd , https://www.jblbbqgrills.com