Proteins from the fractions were extracted by boiling in sample buffer [27 (link)] and separated by SDS-PAGE. Transfer, blocking, and immunodetection of specific proteins were performed largely as previously described [24 (link)]. Dilutions used for the primary antiserum were 1:4,000 for anti-α-tubulin (EMD Millipore, Millerica, MA, USA) and 1:500 for anti-acrosin (Santa Cruz Biotechnology, Dallas, TX, USA), and for the biotin-binding probe was 1:100,000 for NeutrAvidin-HRP (Thermo Fischer Scientific). A 1:5,000 dilution was used for anti-mouse IgG conjugated with HRP (GE Healthcare Life Sciences, Pittsburg, PA, USA). Chemiluminescence was used to detect immunoreactivity.
Anti acrosin
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-acrosin is a laboratory reagent used for research purposes. It is a protein that inhibits the acrosin enzyme, which is involved in the fertilization process of sperm cells. Anti-acrosin is commonly used in various biological and reproductive studies.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse igg conjugated with hrp, by GE Healthcare (1 mentions)
Neutravidin hrp, by Thermo Fisher Scientific (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Pannoramic 250 flash 2 slide scanner, by 3DHISTECH (1 mentions)
Permafluor mounting media, by Thermo Fisher Scientific (1 mentions)
Sodium citrate, by Merck Group (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti acrosin
1
Western Blot Analysis of Sperm Proteins
2
Immunofluorescence Staining of Germ Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Sections were deparaffinized and rehydrated using xylene and ethanol respectively. Epitope retrieval was performed using 10 mM sodium citrate (Sigma-Aldrich) at 95-100°C for 20 min. To block the endogenous peroxidase activity of the tissue, sections were incubated with 3% H 2 O 2 solution for 5 min. Sections were blocked for 1 h with 10% BSA + 0.3% Triton-X in PBS at room temperature and incubated at 4°C overnight with the primary antibodies; anti-Acrosin (1:5, Santa Cruz Biotechnology; Cat. sc-46284), DAZL (1:50, Abcam; Cat. Ab34139). The sections were washed in PBS, 3 times for 10 min each, and then were incubated at room temperature for 1 h with secondary antibodies; Alexa 488 Goat anti-rabbit or donkey anti-goat (both 1:500, Invitrogen Life Technologies). Nuclei were counterstained with Hoechst (1:2000; Invitrogen Life Technologies) for 2 min followed by three 5-min washes with PBS. Slides were protected with PermaFluor mounting media (ThermoFisher Scientific) and then were scanned with a Pannoramic 250 Flash II slide scanner (3DHistech, Budapest, Hungary) at the imaging facility at the Hospital for Sick Children (Toronto, Ontario).
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