Sperm‐cell lysates, SEs‐depleted seminal plasma, and SEs‐protein concentrations were determined using a BCA protein assay kit (P0010S; Beyotime Biotech, Jiangsu, China) according to manufacturer instructions. Subsequently, 30 μg of total protein was heated to 95 °C for 10 min in 1 × DTT‐containing sodium dodecyl sulfate (SDS) sample buffer and separated by 10% SDS‐polyacrylamide gel electrophoresis, followed by transfer onto polyvinylidene fluoride membranes (BioTrace; Bio‐Rad, Hercules, CA, USA). Membranes were blocked with 5% bovine serum albumin for 1 h, and polyclonal antibodies, including anti‐CD81, anti‐HSP‐70, anti‐ALIX, anti‐PHGDH, anti‐LGALS3BP, anti‐SEMG1 (Abclonal, Woburn, MA, USA), anti‐ACTB, anti‐GAPDH (Ray Antibody, Beijing, China), and anti‐Calnexin (Bioworld, Dublin, OH, USA), were used for immunoblotting at 4 °C overnight. Each specific horseradish peroxidase‐conjugated secondary antibody (Ray Antibody) was added accordingly after the membranes were washed in TBS‐Tween solution (TBS‐T) five times for 25 min, and signals were detected by enhanced chemiluminescence (Pierce, Rockford, IL, USA).
Anti calnexin
Manufactured by Bioworld Technology
Sourced in China
Anti-Calnexin is a primary antibody that binds to the calnexin protein, which is an endoplasmic reticulum (ER) resident chaperone involved in the quality control of newly synthesized glycoproteins. This antibody can be used to detect and study the expression and localization of calnexin in various cell and tissue types.
Automatically generated - may contain errors
Lab products found in correlation
Biotrace, by Bio-Rad (5 mentions)
Anti cd81, by ABclonal (5 mentions)
Anti p akt anti alix, by ABclonal (4 mentions)
Anti hmgb1, by ABclonal (4 mentions)
Anti tsg101, by ABclonal (4 mentions)
Anti caspase 3, by ABclonal (4 mentions)
Anti cd63, by ABclonal (4 mentions)
Anti bcl 2, by ABclonal (4 mentions)
Bca protein assay kit, by Beyotime (4 mentions)
Anti akt, by ABclonal (4 mentions)
5 protocols using anti calnexin
1
Seminal Plasma Protein Characterization
2
Protein Quantification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of all the protein we extracted from cells , exosomes and tissues were determined using a BCA protein assay kit ( Beyotime Biotech, Jiangsu, China) according to manufacturer instructions. Subsequently, a certain amount of total protein[20ug-80ug] was heated to 95 °C for 10 min in 1 × DTT-containing sodium dodecyl sulfate (SDS) sample buffer and separated by 10% SDSpolyacrylamide gel electrophoresis, followed by transfer onto polyvinylidene uoride membranes (BioTrace; Bio-Rad, Hercules, CA, USA). Membranes were blocked with 5% bovine serum albumin for 1 h, and polyclonal antibodies, including anti-CD63,anti-CD81, anti-TSG101,anti-AKT,anti-p-AKT anti-ALIX, anti-HMGB1, anti-Caspase-3,anti-Bcl2 (Abclonal, Woburn, MA, USA), anti-ACTB, anti-GAPDH (Ray Antibody, Beijing, China), and anti-Calnexin (Bioworld, Dublin, OH, USA), were used for immunoblotting at 4 °C overnight. Each speci c horseradish peroxidase-conjugated secondary antibody (Ray Antibody) was added accordingly after the membranes were washed in TBS-Tween solution (TBS-T) three times for 30 min, and signals were detected by enhanced chemiluminescence (Pierce, Rockford, IL, USA).
3
Protein Quantification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of all the protein we extracted from cells , exosomes and tissues were determined using a BCA protein assay kit ( Beyotime Biotech, Jiangsu, China) according to manufacturer instructions. Subsequently, a certain amount of total protein[20ug-80ug] was heated to 95 °C for 10 min in 1 × DTT-containing sodium dodecyl sulfate (SDS) sample buffer and separated by 10% SDSpolyacrylamide gel electrophoresis, followed by transfer onto polyvinylidene uoride membranes (BioTrace; Bio-Rad, Hercules, CA, USA). Membranes were blocked with 5% bovine serum albumin for 1 h, and polyclonal antibodies, including anti-CD63,anti-CD81, anti-TSG101,anti-AKT,anti-p-AKT anti-ALIX, anti-HMGB1, anti-Caspase-3,anti-Bcl2 (Abclonal, Woburn, MA, USA), anti-ACTB, anti-GAPDH (Ray Antibody, Beijing, China), and anti-Calnexin (Bioworld, Dublin, OH, USA), were used for immunoblotting at 4 °C overnight. Each speci c horseradish peroxidase-conjugated secondary antibody (Ray Antibody) was added accordingly after the membranes were washed in TBS-Tween solution (TBS-T) three times for 30 min, and signals were detected by enhanced chemiluminescence (Pierce, Rockford, IL, USA).
4
Protein Quantification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of all the protein we extracted from cells , exosomes and tissues were determined using a BCA protein assay kit ( Beyotime Biotech, Jiangsu, China) according to manufacturer instructions. Subsequently, a certain amount of total protein[20ug-80ug] was heated to 95 °C for 10 min in 1 × DTT-containing sodium dodecyl sulfate (SDS) sample buffer and separated by 10% SDSpolyacrylamide gel electrophoresis, followed by transfer onto polyvinylidene uoride membranes (BioTrace; Bio-Rad, Hercules, CA, USA). Membranes were blocked with 5% bovine serum albumin for 1 h, and polyclonal antibodies, including anti-CD63,anti-CD81, anti-TSG101,anti-AKT,anti-p-AKT anti-ALIX, anti-HMGB1, anti-Caspase-3,anti-Bcl2 (Abclonal, Woburn, MA, USA), anti-ACTB, anti-GAPDH (Ray Antibody, Beijing, China), and anti-Calnexin (Bioworld, Dublin, OH, USA), were used for immunoblotting at 4 °C overnight. Each speci c horseradish peroxidase-conjugated secondary antibody (Ray Antibody) was added accordingly after the membranes were washed in TBS-Tween solution (TBS-T) three times for 30 min, and signals were detected by enhanced chemiluminescence (Pierce, Rockford, IL, USA).
5
Protein Quantification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of all the protein we extracted from cells , exosomes and tissues were determined using a BCA protein assay kit ( Beyotime Biotech, Jiangsu, China) according to manufacturer instructions. Subsequently, a certain amount of total protein[20ug-80ug] was heated to 95 °C for 10 min in 1 × DTT-containing sodium dodecyl sulfate (SDS) sample buffer and separated by 10% SDSpolyacrylamide gel electrophoresis, followed by transfer onto polyvinylidene uoride membranes (BioTrace; Bio-Rad, Hercules, CA, USA). Membranes were blocked with 5% bovine serum albumin for 1 h, and polyclonal antibodies, including anti-CD63,anti-CD81, anti-TSG101,anti-AKT,anti-p-AKT anti-ALIX, anti-HMGB1, anti-Caspase-3,anti-Bcl2 (Abclonal, Woburn, MA, USA), anti-ACTB, anti-GAPDH (Ray Antibody, Beijing, China), and anti-Calnexin (Bioworld, Dublin, OH, USA), were used for immunoblotting at 4 °C overnight. Each speci c horseradish peroxidase-conjugated secondary antibody (Ray Antibody) was added accordingly after the membranes were washed in TBS-Tween solution (TBS-T) three times for 30 min, and signals were detected by enhanced chemiluminescence (Pierce, Rockford, IL, USA).
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