How to prepare Tissue Homogenate samples?
2025-05-06
Tissue Homogenate
1) Rinse the tissue sample with PBS (0.01M, pH 7.4) to wash away the residual blood or impurities on the surface of the tissue.
2) Weigh the tissue block, record the weight, and then cut it into small pieces. The pieces should be as small as possible to ensure
more thorough homogenization.
3) Add the tissue to pre-cooled PBS at a certain ratio (add the protease inhibitor immediately before use, and refer to the instructions
for use of the inhibitor for the addition ratio). Homogenize the tissue while placing it on ice or in an ice bath. (Generally, homogenize
according to the ratio of tissue weight to PBS volume of 1:9. For example, 1g of tissue sample corresponds to 9mL of PBS. The specific
volume can be adjusted appropriately according to the experimental needs. When calculating the sample concentration after detection,
the corresponding dilution factor should be multiplied.)
4) Aspirate the homogenate into a centrifuge tube, centrifuge at 4°C and 5000×g for 5-10 minutes. Take the supernatant and store it at
-20°C or -80°C. Avoid repeated freezing and thawing.
more thorough homogenization.
3) Add the tissue to pre-cooled PBS at a certain ratio (add the protease inhibitor immediately before use, and refer to the instructions
for use of the inhibitor for the addition ratio). Homogenize the tissue while placing it on ice or in an ice bath. (Generally, homogenize
according to the ratio of tissue weight to PBS volume of 1:9. For example, 1g of tissue sample corresponds to 9mL of PBS. The specific
volume can be adjusted appropriately according to the experimental needs. When calculating the sample concentration after detection,
the corresponding dilution factor should be multiplied.)
4) Aspirate the homogenate into a centrifuge tube, centrifuge at 4°C and 5000×g for 5-10 minutes. Take the supernatant and store it at
-20°C or -80°C. Avoid repeated freezing and thawing.
Should we add a protease and phosphatase inhibitor cocktail during homogenization?
If the sample needs to be stored for a long time, it is necessary to add the inhibitor cocktail. If the detection is
carried out immediately after homogenization, the protease inhibitor can be omitted.
If the sample needs to be stored for a long time, it is necessary to add the inhibitor cocktail. If the detection is
carried out immediately after homogenization, the protease inhibitor can be omitted.
Homogenize at a ratio of tissue weight to PBS volume of 1:9. For example, 1 g of tissue sample corresponds to 9 mL of PBS. The specific
volume can be appropriately adjusted according to experimental requirements. When calculating the sample concentration after detection,
the corresponding dilution factor should be multiplied.
volume can be appropriately adjusted according to experimental requirements. When calculating the sample concentration after detection,
the corresponding dilution factor should be multiplied.
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