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Method for extracting protein from frozen cow semen

A technology for freezing semen and sperm, applied in the field of bioengineering, can solve problems such as unclear mechanism of action and impact on semen quality

Inactive Publication Date: 2013-05-22
NORTHWEST A & F UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Artificial insemination technology is one of the most widely used technologies in the cattle industry. Although it has achieved good results, the general research only involves the effect of different diluents on the quality of semen freezing.
There are relatively few studies on the mechanism of improving sperm motility by different components in the diluent, and the related genes, proteins, and their interaction and expression signaling pathways that affect the motility of sperm after freezing. Because the mechanism of action is not clear, it will affect the quality of semen further improvement of

Method used

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  • Method for extracting protein from frozen cow semen
  • Method for extracting protein from frozen cow semen
  • Method for extracting protein from frozen cow semen

Examples

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Embodiment 1

[0016] 1) Thaw the frozen semen and transfer it to a centrifuge tube, centrifuge at 4 °C and 1000×g for 5 min, and discard the supernatant;

[0017] 2) Add 1 ml of PBS pre-cooled to 4 ℃ for washing, centrifuge at 4 ℃, 1000×g for 5 min, repeat 3 times;

[0018] 3) Add 200 μl of protein lysate (solarbio, RIPA tissue / cell lysate) to the above precipitate, and vortex with a vortex instrument (2800 r / min) to fully dissolve the sample;

[0019] 4) After standing for 5 minutes, centrifuge at 4 °C and 12000 × g for 30 minutes, and the supernatant is the sperm protein solution.

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Abstract

The invention discloses a method for extracting protein from frozen cow semen. The method comprises the following steps of: unfreezing frozen semen and transferring to a centrifugal tube, centrifuging the unfrozen semen for 5 minutes at 4 DEG C and 1000*g, discarding the liquid supernatant; (2), adding 1ml of PBS (Phosphate Buffer Solution) which is pre-cooled to 4 DEG C, centrifuging for 5 minutes at 4 DEG C and 1000*g, repeating the process for three times; (3), adding 200 microliters of protein lysis solution to the precipitate and sufficiently dissolving the sample by volution; (4), stewing the solution for 5 minutes and centrifuging the solution for 30 minutes at 4 DEG C and 12000*g, wherein the obtained liquid supernatant is protein solution. The method for extracting protein from frozen cow semen disclosed by the invention is simple to operate, capable of laying a foundation for researching the semen freezing and protecting mechanism by molecular means in the future and capable of further improving the activity of the frozen semen.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a bovine frozen semen sperm quality assessment technology, in particular to a method for extracting protein from bovine frozen sperm. [0002] Background technique [0003] The cryopreservation of domestic animal semen has been produced with the widespread application of artificial insemination technology since the 20th century. The speed of sperm metabolism is reduced or inhibited by ultra-low temperature, so that the life of sperm is preserved in a relatively static state to prolong the time of in vitro survival. The application of semen cryopreservation not only solves the problem of long-term storage of semen, facilitates cooperation and exchanges at home and abroad, reduces the cost of artificial insemination, but also facilitates the efficient use of genetic resources of excellent breeding males, accelerates genetic progress, accelerates breed improvement, and improve...

Claims

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Application Information

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IPC IPC(8): C07K1/14
Inventor 胡建宏王红李青旺王鹏洪洁赟王艳风侯放亮马国际贾永宏
Owner NORTHWEST A & F UNIV
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