Total protein was extracted using protein lysis buffer with ground central inflorescences. 5× SDS loading buffer was added to the supernatants and the lysates were boiled at 95 °C for 6 min. Samples were separated by SDS-PAGE using 10% acrylamide gels and electroblotted to nitrocellulose (NC) membranes (Abm, B500) and incubated in anti-FLAG, -MYC, -GFP, -UBQ11 (Agrisera, AS08 307A, 1:5000 dilution), -DMC1 (1:500 dilution)65 (link), -β-Tubulin (Abmart, M20005, 1:2000 dilution), and -HSC70 (ENZO, ADI-SPA-818-F, 1:2000 dilution) antibodies. Proteins were detected using corresponding antibodies. Bands were visualized with a Clinx-3400 chemiluminescence imaging system. Goat anti-mouse IgG HRP-conjugated antibody (GNI, GNI9310-M, 1:2000 dilution) and goat anti-rabbit IgG HRP-conjugated antibody (GNI, GNI9310-R, 1:2000 dilution) were used as the secondary antibody. Source data of the most important blots are provided.
As08 307a
Manufactured by Agrisera
The AS08 307A is a laboratory equipment product offered by Agrisera. It serves as a core component in various research and analytical applications. The product's primary function is to perform a specific set of tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
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Lab products found in correlation
2 protocols using as08 307a
1
Protein Extraction and Western Blotting
2
Western Blot Analysis of Protein Samples
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Protein samples were boiled for 5 min. After a 2-min centrifugation at 13,000 rpm, the supernatants were loaded on a 10% SDS-PAGE gel. Rabbit anti-RHD3CT antibody (PHY0765S; PhytoAB; 1:10,000 dilution), anti-AGB1 antibody (PHY0956S; PhytoAB; 1:1000 dilution), anti-LPAT2 antibody (PHY0873S; PhytoAB; 1:1000 dilution), anti-actin antibody (AS13 2640; Agrisera; 1:4000 dilution), anti-GST antibody (AS17 4147; Agrisera; 1:2000 dilution), and anti-ubiquitin antibody (AS08 307A; Agrisera; 1:1000 dilution) were used for the protein gel blot. Mouse anti-tubulin (T6074; Sigma-Aldrich; 1:4000) and goat anti-mouse (A4914-1ML; Sigma-Aldrich; 1:10,000) were used as the loading control of the protein gel blot. For the blotting after immunoprecipitation, a rabbit anti-GFP antibody (ab32146; Abcam) at 1:5000 dilution or a rabbit anti-RFP (ab34771; Abcam) at 1:2000 dilution was used. The secondary anti-rabbit IgG-peroxidase (Sigma-Aldrich) was used at 1:5000 dilution. The intensity of bands was quantified relatively to the reference band by ImageJ.
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