Special Topics on column protein extraction| Abbkine ExKine™ Pro Series Kits

Disadvantages of conventional RIPA lysis buffer protein extraction

The conventional RIPA lysis buffer is easy to produce insoluble components for protein extraction, resulting in random loss of proteins, incomplete protein obtained, affected protein activity, and low protein purity, thus limiting the applicability of protein samples obtained for downstream experiments.

Figure 1. Abbkine Protein Extraction Kit (Column method)

Figure 2. Comparison of solubility of Abbkine protein extraction kit (column method) lysates and conventional RIPA lysis buffer.

(Sample: 293 cells)

Compared with the conventional RIPA lysis buffer liquid phase of various strengths, the lysis buffer of Abbkine protein extraction kit (column method) showed higher protein solubility, and no soluble components were produced, which could solve the problem of total protein loss in solution extraction.

Advantages of Abbkine protein extraction kit (column method)

ExKine™ Pro Total Protein Extraction Kit for Animal Cultured Cells/Tissues

• Simple and fast: it can be completed within 1-7 min;
• High protein yield: no protein loss, 50 mg animal tissue, protein yield up to 3-20 mg/mL;
• Mild pyrolysis conditions: without ultrasonic broken, boil, repeated freezing and thawing and other harsh conditions;
• High solubility: fat-soluble and water-soluble protein extraction, solubility than RIPA;
• Choice: provide two (natural/degeneration) pyrolysis liquid, meet the demand of extraction degeneration and non denatured protein;
• Good compatibility: compatible with more downstream experiment, the protein is suitable for SDS-PAGE, WB, IP, ELISA and enzyme activity detection experiments.
• ExKine™ Pro Total Protein Extraction Kit for Plant Tissues
• Convenient: just need 8 minutes;
• Many uses: for samples to be complete – plant tissue soft leaves, seeds, stems, roots;
• High protein yield: from 50-200 mg of plant tissue, total protein yield can be up to 2-8 mg/mL;
• Operation simple and safe, without liquid nitrogen grinding and optimize the pyrolysis buffer more efficient protein;
• Good compatibility: the protein can be used for SDS-PAGE, WB, IP, ELISA, downstream of the enzyme activity detection experiments.

Figure 3. SDS-PAGA of Animal protein obtained by Abbkine ExKine™ Pro Total Protein Extraction Kit for Animal Cultured Cells/Tissues (column method) with the same column protein extraction kit from other brands (Abbkine KTP3007)。

Note: 1-RIPA; 2-Abbkine denatured cleavage; 3-A company denaturing and cracking; Denaturing and cracking of 4-B company; 5-C company denaturing and cracking; 6-Abbkine nondenatured cleavage; 7-C non-denatured cracking; M-Marker. The sample was mouse liver with the loading amount of 4 uL.

The Abbkine protein extraction kit has a wider total protein expression profile than the same column protein extraction kit from other brands.

In order to verify the integrity of membrane proteins and nuclear proteins extracted from total proteins by Abbkine total protein extraction kit, proteins in mouse liver cells and 293 cells were incubated with Histone H3 antibody and ATPase, respectively, and compared with proteins extracted by D Company’s kit. The results are shown as follows:

Figure 4. Integrity verification of membrane proteins and nuclear proteins in proteins extracted from animal protein extraction products using column method from different companies.

Note: 1- Add 5 μL protein extract from D Company’s kit; 2- Add 2.5 μL protein extract extracted by D Company’s kit; 3- Add 5 μL protein extract from Abbkine kit; 4- Add 2.5 μL protein extract from Abbkine kit.

The results showed that Abbkine column protein extraction kit could obtain clear background protein bands in both cell and tissue samples. The extracted nucleoprotein signal is strong, and the effect of membrane protein extraction is significantly better than that of other brands, which solves the problem that membrane protein is easily lost in the process of total protein extraction.

Figure 5. Total proteins were extracted from different plants (Arabidopsis, soybean) for verification.

Note: The left lane is Arabidopsis and the right lane is soybean.

The results showed that ExKine™ Pro plant tissue total protein extraction kit (column method) was highly effective in all plant samples.

Abbkine ExKine™ Pro Column method protein extraction series makes your protein qualitative and quantitative tests more accurate, easier phosphorylation detection, and easy access to hard-to-enrich membrane proteins. In addition to bring you 7 minutes of super fast protein extraction operation experience, for your protein experiment escort!

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