Human SP-10/ACRV1 HEK293 Overexpression Lysate

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Human SP-10/ACRV1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human SP-10/ACRV1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SP-10/ACRV1 protein (Cat: 11789-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the mature form of human ACRV1 isoform 1 (P26436-1) (Gln 22-Ile 265) was fused with a polyhistidine tag at the C-terminus and a signal peptide at the N-terminus.
Molecule Mass
The recombinant human ACRV1 consists of 255 amino acids and predictes a molecular mass of 27.3 kDa. In SDS-PAGE under reducing conditions, rhACRV1 migrates as an approximately 38 kDa band due to glycosylation.

Human SP-10/ACRV1 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human SP-10/ACRV1 HEK293 Overexpression Lysate: Alternative Names

Human D11S4365 Overexpression Lysate; Human SP-10 Overexpression Lysate; Human SPACA2 Overexpression Lysate

SP-10/ACRV1 Background Information

Acrosomal protein SP-1, also known as Acrosomal vesicle protein 1 and ACRV1, is a testis-specific, differentiation antigen, that arises within the acrosomal vesicle during spermatogenesis, and is associated with the acrosomal membranes and matrix of mature sperm. Regulation of cell type-specific gene transcription is central to cellular differentiation and development. During spermatogenesis, a number of testis-specific genes are expressed in a precise spatiotemporal order. The longest transcript of ACRV1 / SP-1 is the most abundant, comprising 53-72% of the total acrosomal vesicle protein 1 messages; the second largest transcript comprises 15-32%; the third and the fourth largest transcripts account for 3.4-8.3% and 8.7-12.5%, respectively; and the remaining transcripts combined account for < 1% of the total acrosomal vesicle protein 1 message. ACRV1 / SP-1 is a testis-specific acrosomal protein that has been detected in several species including humans. It may be involved in sperm-zona binding or penetration, and it is a potential contraceptive vaccine immunogen for humans. ACRV1 / SP-1 may be involved in sperm-zona binding or penetration. It is also a intra-acrosomal protein that is considered to be a vaccine candidate for immunocontraception.
Full Name
acrosomal vesicle protein 1
References
  • Wright RM, et al.,1990, Biol Reprod. 42 (4): 693-701.
  • Kurth,B.E. et al., 1993, Anat Rec. 236 (4):619-25.
  • Golden WL, et al.,1994, Genomics.18 (2): 446-9. 
  • Foster,J.A. et al., 1994, Biol Reprod. 51 (6):1222-31.
  • Abhyankar,M.M. et al., 2007, J Biol Chem. 282 (50): 36143-54.
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