Samples were diluted in SDS-PAGE Laemmli loading buffer (Bio-Rad) and electrophoresed on a 4–20% Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred to nitrocellulose membranes using a Trans-Blot Turbo transfer system. Blots were blocked in 5% milk/PBST (1× PBS [pH 7.4], 0.1% Tween 20) and then washed with PBST. In-house-made primary antibody, CR3022 as previously described14 (link),70 (approximate concentration 0.8–1.3 mg/mL), was added at a 1:10,000 dilution in PBST. Blots were washed with PBST, and secondary rabbit anti-human IgG H&L HRP (abcam ab6759) was added at 1:10,000 in PBST. Blots were developed using Pierce ECL substrate and imaged using a GE Amersham imager 600.
Sds page laemmli loading buffer
Manufactured by Bio-Rad
Sourced in United States
SDS-PAGE Laemmli loading buffer is a solution used to prepare protein samples for separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). It contains the anionic detergent SDS, which denatures and solubilizes proteins, as well as a reducing agent, glycerol, and tracking dyes to facilitate sample loading and visualization during electrophoresis.
Automatically generated - may contain errors
6 protocols using sds page laemmli loading buffer
1
Immunoblotting Assay for SARS-CoV-2 Antibody Detection
2
SARS-CoV-2 Antibody Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expi293F culture supernatants were collected
3 days after transfection,
harvested via spinning at 7000g for 15 min, and filtered
through a 0.22 μm filter. Samples were diluted in SDS-PAGE Laemmli
loading buffer (Bio-Rad), boiled at 95 °C, and run on a 4–20%
Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred
to nitrocellulose membranes using a Trans-Blot Turbo transfer system.
Blots were blocked in 5% milk/PBST (1× PBS [pH 7.4], 0.1% Tween
20) and then washed with PBST. In-house-made primary antibodies CR3022,
COVA2-15, and CB6 (approximate concentrations 0.8–1.3 mg/mL)
were added at a 1:10 000 dilution in PBST. Blots were washed
with PBST, and secondary rabbit antihuman IgG H&L HRP (abcam ab6759)
was added at 1:10 000 in PBST. Blots were developed using Pierce
ECL substrate and imaged using a GE Amersham imager 600.
3 days after transfection,
harvested via spinning at 7000g for 15 min, and filtered
through a 0.22 μm filter. Samples were diluted in SDS-PAGE Laemmli
loading buffer (Bio-Rad), boiled at 95 °C, and run on a 4–20%
Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred
to nitrocellulose membranes using a Trans-Blot Turbo transfer system.
Blots were blocked in 5% milk/PBST (1× PBS [pH 7.4], 0.1% Tween
20) and then washed with PBST. In-house-made primary antibodies CR3022,
COVA2-15, and CB6 (approximate concentrations 0.8–1.3 mg/mL)
were added at a 1:10 000 dilution in PBST. Blots were washed
with PBST, and secondary rabbit antihuman IgG H&L HRP (abcam ab6759)
was added at 1:10 000 in PBST. Blots were developed using Pierce
ECL substrate and imaged using a GE Amersham imager 600.
3
Western Blot Analysis of COVID-19 Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Expi293F culture supernatants were collected 3 days after transfection, harvested via spinning at 7,000 × g for 15 minutes, and filtered through a 0.22-μm filter. Samples were diluted in SDS-PAGE Laemmli loading buffer (Bio-Rad), boiled at 95 °C, and run on a 4–20% Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred to nitrocellulose membranes using a Trans-Blot Turbo transfer system. Blots were blocked in 5% milk / PBST (1X PBS [pH 7.4], 0.1% Tween 20) and then washed with PBST. In-house made primary antibodies CR3022, COVA2–15, and CB6 (approximate concentrations 0.8 – 1.3 mg/mL) were added at a 1:10,000 dilution in PBST. Blots were washed with PBST and secondary rabbit anti-human IgG H&L HRP (abcam ab6759) was added at 1:10,000 in PBST. Blots were developed using Pierce ECL substrate and imaged using a GE Amersham Imager 600.
4
Deglycosylated Aortic Valve ECM Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Alkylated and deglycosylated aortic valve ECM samples (procedure described above) were slowly thawed and kept on ice. SDS-PAGE Laemmli Loading buffer (BioRad, USA) with β-mercaptoethanol was added to the samples, and the mixtures were boiled at 100 °C for 5 min. The samples were resolved on SDS-PAGE 4–20% gradient gels and transferred to a polyvinylidene difluoride (PVDF) membrane. The membranes were blocked in 5% skimmed milk in Tris-buffered saline + 0.05% Tween20 (TBS-T; pH 7.6) and, after that, hybridized overnight with primary antibodies of interest. All washing steps were made with TBS-T except for the last step that was made with TBS. Proteins were visualized using Clarity Western ECL Substrate (BioRad, USA) and scanned using BioRad Chemi Doc Universal Hood II using the QuantiOne program.
5
Immunoblotting Assay for SARS-CoV-2 Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were diluted in SDS-PAGE Laemmli loading buffer (Bio-Rad) and electrophoresed on a 4–20% Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred to nitrocellulose membranes using a Trans-Blot Turbo transfer system. Blots were blocked in 5% milk/PBST (1× PBS [pH 7.4], 0.1% Tween 20) and then washed with PBST. In-house-made primary antibody, CR3022 as previously described14 (link),70 (approximate concentration 0.8–1.3 mg/mL), was added at a 1:10,000 dilution in PBST. Blots were washed with PBST, and secondary rabbit anti-human IgG H&L HRP (abcam ab6759) was added at 1:10,000 in PBST. Blots were developed using Pierce ECL substrate and imaged using a GE Amersham imager 600.
6
Immunoblotting Assay for SARS-CoV-2 Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were diluted in SDS-PAGE Laemmli loading buffer (Bio-Rad) and electrophoresed on a 4–20% Mini-PROTEAN TGX protein gel (Bio-Rad). Proteins were transferred to nitrocellulose membranes using a Trans-Blot Turbo transfer system. Blots were blocked in 5% milk/PBST (1× PBS [pH 7.4], 0.1% Tween 20) and then washed with PBST. In-house-made primary antibody, CR3022 as previously described14 (link),70 (approximate concentration 0.8–1.3 mg/mL), was added at a 1:10,000 dilution in PBST. Blots were washed with PBST, and secondary rabbit anti-human IgG H&L HRP (abcam ab6759) was added at 1:10,000 in PBST. Blots were developed using Pierce ECL substrate and imaged using a GE Amersham imager 600.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!