Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1 × PBS (pH 7.4) and lysing them with 1 × SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (Amersham) overnight (ON) using a Bio-Rad Trans-Blot. EVs were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated ON at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500-diluted anti-β-actin AC-74 mAb from Sigma, 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz, and 1:1000 diluted anti-flag M2 mAb from Sigma. Filters were analyzed by a Chemi-Doc apparatus, Bio-Rad, and relevant signals were quantified by Image Lab software version 6.1 (Bio-Rad, Hercules, CA, USA).
Anti alix h 270 polyclonal abs
Manufactured by Santa Cruz Biotechnology
Sourced in United Kingdom, United States
Anti-Alix H-270 polyclonal Abs is a laboratory reagent produced by Santa Cruz Biotechnology. It is a polyclonal antibody that targets the Alix protein.
Automatically generated - may contain errors
Lab products found in correlation
Trans blot, by Bio-Rad (6 mentions)
Anti β actin ac 74 mab, by Merck Group (4 mentions)
Pore size nitrocellulose membrane, by Cytiva (4 mentions)
Anti actin ac 74 mab, by Merck Group (3 mentions)
Anti flag m2 mab, by Merck Group (2 mentions)
Pore size nitrocellulose membrane, by GE Healthcare (2 mentions)
Chemidoc apparatus, by Bio-Rad (1 mentions)
Pore size nitrocellulose membrane, by Bio-Rad (1 mentions)
Image lab software version 6, by Bio-Rad (1 mentions)
Transblot device, by Bio-Rad (1 mentions)
Chemidoc imager, by Bio-Rad (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
7 protocols using anti alix h 270 polyclonal abs
1
Comprehensive Western Blot Analysis of Cells and EVs
2
Western Blot Analysis of Cell Lysates and EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1× PBS (pH 7.4) and lysing them with 1× SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (GE Healthcare Europe GmbH, Milan, Italy ) overnight using a Bio-Rad (Hercules, CA, USA) trans-blot device. For Western blot analysis of EVs, they were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500-diluted anti-β-actin AC-74 mAb from Sigma (St. Louis, MI, USA), 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz (Dallas, TX, USA), and 1:1000 diluted 6AT18 anti-6×His-tag mAb (Sigma).
3
Western Blot Analysis of Cell Lysates and EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1× PBS (pH 7.4) and lysing them with 1 × SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (GE Healthcare Europe GmbH, Milan, Italy) overnight using a Bio-Rad (Hercules, CA, USA) Trans-Blot. For western blot analysis of EVs, they were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500-diluted anti-β-actin AC-74 mAb from Sigma (St. Louis, MI, USA), and 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz (Dallas, TX, USA).
4
Exosomal and Cellular Protein Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and exosomes were carried out as described [4 (link)] after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, Western blot analysis on cell lysates was performed by washing cells twice with 1 × PBS (pH 7.4) and lysing them with 1 × SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45-μM pore size nitrocellulose membrane (Amersham) overnight using a Bio-Rad (Hercules, CA, USA) Trans-Blot. For Western blot analysis of exosomes, they were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500-diluted anti-β-actin AC-74 mAb from Sigma (St. Louis, MO, USA), and 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz (Heidelberg, Germany). Densitometry analysis was carried out with Bio-Rad Image Lab software of a ChemiDoc imager.
5
Western Blot Analysis of Cell Lysates and EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1×PBS (pH 7.4) and lysing them with 1× SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (Amersham) overnight using a Bio-Rad Trans-Blot. EVs were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1,000diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500-diluted anti--actin AC-74 mAb from Sigma, 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz, and 1:1,000 diluted anti-flag M2 mAb from Sigma.
6
Western Blot Analysis of Cell Lysates and EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1×PBS (pH 7.4) and lysing them with 1x SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (Amersham) overnight using a Bio-Rad Trans-Blot. For western blot analysis of EVs, they were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1,000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500diluted anti--actin AC-74 mAb from Sigma, and 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz.
7
Western Blot Analysis of Cell Lysates and EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analyses of both cell lysates and EVs were carried out after resolving samples in 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In brief, the analysis on cell lysates was performed by washing cells twice with 1×PBS (pH 7.4) and lysing them with 1x SDS-PAGE sample buffer. Samples were resolved by SDS-PAGE and transferred by electroblotting on a 0.45 μM pore size nitrocellulose membrane (Amersham) overnight using a Bio-Rad Trans-Blot. For western blot analysis of EVs, they were lysed and analyzed as described for cell lysates. For immunoassays, membranes were blocked with 5% non-fat dry milk in PBS containing 0.1% Triton X-100 for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies diluted in PBS containing 0.1% Triton X-100. Filters were revealed using 1:1,000-diluted sheep anti-Nef antiserum ARP 444 (MHRC, London, UK), 1:500diluted anti--actin AC-74 mAb from Sigma, and 1:500 diluted anti-Alix H-270 polyclonal Abs from Santa Cruz.
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!