Cells or extracellular vesicles fraction were lysed in RIPA buffer (Santa Cruz, USA) and cleared lysates were collected by centrifugation at 12,000 × g for 10 min at 4°C. Lysates were boiled in 5 × gel loading buffer and separated by SDS‐PAGE gels. Proteins were transferred onto PVDF membranes (MERCK‐millipore) and detected with appropriate primary antibodies CD9 (Sigma, # SAB4503606), TSG101 (#49270, Signalway Antibody), GAPDH (Proteintech, #60004‐1), Annexin A5 (Abcam, #ab171870), Annexin A1 (Cell Signaling Technology, #32934), GM130 (Cell Signaling Technology, #12480), and Calnexin (Cell Signaling Technology, #2679) at 4˚C overnight. The HRP‐conjugated goat antimouse or rabbit IgG (Southern Biotech) was used and exposed using the chemiluminescence (ECL) (Millipore, #WBLUC0100). The band intensities were quantified by ImageJ software (NIH).
Hrp conjugated goat antimouse or rabbit igg
Manufactured by Southern Biotech
The HRP-conjugated goat anti-mouse or rabbit IgG is a secondary antibody used in various immunoassay techniques. It binds to the primary antibody, which is raised against a specific target antigen, and is conjugated with horseradish peroxidase (HRP) enzyme. The HRP enzyme can catalyze a color-producing reaction, allowing for the detection and quantification of the target antigen.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (2 mentions)
Ripa buffer, by Santa Cruz Biotechnology (2 mentions)
Calnexin, by Cell Signaling Technology (1 mentions)
Gm130, by Cell Signaling Technology (1 mentions)
Annexin a1, by Cell Signaling Technology (1 mentions)
Tsg101, by Signalway Antibody (1 mentions)
Gapdh, by Proteintech (1 mentions)
Flag tag m2, by Merck Group (1 mentions)
Sars cov 2 rbd, by Sino Biological (1 mentions)
Hace 2, by Abcam (1 mentions)
Gm130, by Abcam (1 mentions)
Calnexin, by Abcam (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
2 protocols using hrp conjugated goat antimouse or rabbit igg
1
Comprehensive Extracellular Vesicle Protein Analysis
2
Western Blot Analysis of SARS-CoV-2 RBD and EV Markers
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Cells, purified EVs or tissues were lysed with RIPA buffer (Santa Cruz,USA) and cleared lysate was collected by centrifugation for protein separation on SDS-polyacrylamide gel(10%), followed by transfer onto PVDF membranes(Millipore), blocked with 5% nonfat dry milk in TBST and reacted with primary antibodies recognizing FLAG M2 tag(Sigma,1:2000), SARS-CoV-2 RBD(Sinobiological,1:500), CD9(Abcam,1:2000), Alix(Cell Signaling Technology,1:1000), GM130(Abcam,1:1000), Calnexin(Abcam,1:1000), hACE2(Abcam,# ab108209,only react with human species,1:2000) and GAPDH(Santa Cruz,1:2000). Blots were incubated with HRP-conjugated goat anti-mouse or rabbit IgG (Southern Biotech,1:5000) for 1 h at room temperature before imaging.
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