Cell lysates were prepared by washing cells in cold PBS followed by addition of SDS-PAGE sample buffer containing Protease Inhibitor Cocktail (SIGMA Cat#P2714–1BTL). Cells were transferred to a 1.5ml tube, sonicated, and heated at 95°C for 5 min. Proteins were separated by electrophoresis in NuPAGE 10% Bis-Tris gels and transferred to PVDF membranes (Milipore- Immobilon, FL, Cat #IPFL00010- pore size: 0.45μm) by electroblotting. Membranes were blocked in PBS+ 5% milk powder and incubated with primary antibodies and anti-SEPT9 rabbit antibody (Proteintech Cat#10769–1-AP) and α-TUBULIN mouse antibody (Sigma Cat #T9026) overnight at 4°C. The other antibodies used were MMP3 rabbit antibody (Arigo Biolaboratories Cat#ARG55262), CD9 mouse antibody (Proteintech Cat#60232–1-Ig), or Actin mouse antibody (Proteintech Cat#66009–1-Ig). After three washes in PBS-T, membranes were incubated with secondary antibodies (Mouse 680 and Rabbit 800 [LI-COR Biosciences]) for one hour at room temperature, washed three times in PBS-T, and scanned using a high-sensitivity Odyssey Infrared Imaging System (Li-COR Biosciences). α-TUBULIN was used as a loading control. ImageJ was used for quantification of bands intensities.
Cd9 mouse antibody
Manufactured by Proteintech
The CD9 mouse antibody is a laboratory reagent used for the detection and analysis of the CD9 protein in biological samples. CD9 is a member of the tetraspanin family of proteins and is commonly expressed on the surface of various cell types, including immune cells and exosomes. This antibody can be used in techniques such as flow cytometry, immunoprecipitation, and Western blotting to study the expression and localization of CD9 in research applications.
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2 protocols using cd9 mouse antibody
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Western Blot Analysis of Cell Lysates
2
Western Blot Analysis of Cell Lysates
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Cell lysates were prepared by washing cells in cold PBS followed by addition of SDS-PAGE sample buffer containing Protease Inhibitor Cocktail (SIGMA Cat#P2714–1BTL). Cells were transferred to a 1.5ml tube, sonicated, and heated at 95°C for 5 min. Proteins were separated by electrophoresis in NuPAGE 10% Bis-Tris gels and transferred to PVDF membranes (Milipore- Immobilon, FL, Cat #IPFL00010- pore size: 0.45μm) by electroblotting. Membranes were blocked in PBS+ 5% milk powder and incubated with primary antibodies and anti-SEPT9 rabbit antibody (Proteintech Cat#10769–1-AP) and α-TUBULIN mouse antibody (Sigma Cat #T9026) overnight at 4°C. The other antibodies used were MMP3 rabbit antibody (Arigo Biolaboratories Cat#ARG55262), CD9 mouse antibody (Proteintech Cat#60232–1-Ig), or Actin mouse antibody (Proteintech Cat#66009–1-Ig). After three washes in PBS-T, membranes were incubated with secondary antibodies (Mouse 680 and Rabbit 800 [LI-COR Biosciences]) for one hour at room temperature, washed three times in PBS-T, and scanned using a high-sensitivity Odyssey Infrared Imaging System (Li-COR Biosciences). α-TUBULIN was used as a loading control. ImageJ was used for quantification of bands intensities.
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