准备和存储
推荐的实验流程
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
The cAMP-dependent Protein Kinase (PKA) is compartmentalized within the cell. To maintain the localization of PKA, the regulatory subunits interact with specific anchoring proteins. Several proteins have been identified as PKA anchoring proteins and form a family named AKAP (A-Kinase Anchor Proteins). Fifteen of the AKAP proteins contain a consensus binding motif that allows interaction with the type II regulatory subunit (RII) of the PKA holoenzyme. In addition, three other AKAPs (D-AKAP1, D-AKAP2, and fsc1/AKAP82) can associate with the type I regulatory subunit (RI) of the PKA holoenzyme. AKAP82 was isolated as a component of the mouse sperm fibrous sheath. It is a dual specificity AKAP that contains an RII-binding domain (domain A; amino acids 219 to 232) and an RI-binding domain (domain B; amino acids 335-344). In mouse, pro-AKAP82 is synthesized as a 97 kDa precursor that is transported to the flagellum where proteolytic cleavage of the N-terminal 179 amino acids produces AKAP82. Assembly of AKAP82 into the fibrous sheath surrounding the axoneme of the sperm flagellum is thought to tether PKA close to the axoneme where it can regulate flagellar motility.
研发参考 (4)
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Edwards AS, Scott JD. A-kinase anchoring proteins: protein kinase A and beyond. Curr Opin Immunol. 2000; 12(2):217-221. (Biology). 查看参考
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Fulcher KD, Mori C, Welch JE, O'Brien DA, Klapper DG, Eddy EM. Characterization of Fsc1 cDNA for a mouse sperm fibrous sheath component. Biol Reprod. 1995; 52(1):41-49. (Biology). 查看参考
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Miki K, Eddy EM. Identification of tethering domains for protein kinase A type Ialpha regulatory subunits on sperm fibrous sheath protein FSC1. J Biol Chem. 1998; 273(51):34384-34390. (Biology). 查看参考
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Turner RM, Johnson LR, Haig-Ladewig L, Gerton GL, Moss SB. An X-linked gene encodes a major human sperm fibrous sheath protein, hAKAP82. Genomic organization, protein kinase A-RII binding, and distribution of the precursor in the sperm tail. J Biol Chem. 1998; 273(48):32135-32141. (Biology). 查看参考
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