The interaction between USP10 and KLF4 was detected by Co-IP. BMSCs were lysed using Co-IP lysate to extract proteins. Protein supernatants were incubated with Normal rabbit IgG or USP10 (AB70895, Abcam), and SKP2 (AB214666, Abcam) antibodies at 4℃ overnight with rotation. The Co-IP lysate was then added to Protein A/G agarose beads, mixed, and centrifuged to collect the precipitate. Following antibody incubation, the cell lysate was fully incubated with agarose beads for 2 h at 4℃ to allow antibody coupling to the beads. The agarose beads were washed using Co-IP lysate buffer, and WB subsequently analyzed the coupling product.
Ab70895
Manufactured by Abcam
AB70895 is a laboratory equipment product offered by Abcam. It is a device designed for a specific scientific purpose, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Ab214666, by Abcam (2 mentions)
Alexa fluor donkey anti mouse 488 nm, by Thermo Fisher Scientific (1 mentions)
Mabe343, by Merck Group (1 mentions)
Alexa fluor goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Ab40693, by Merck Group (1 mentions)
Alexafluor donkey anti rabbit 488nm, by Thermo Fisher Scientific (1 mentions)
Ab18197, by Abcam (1 mentions)
Hrp conjugated goat anti rabbit, by Bio-Rad (1 mentions)
Hrp conjugated goat anti mouse, by Bio-Rad (1 mentions)
3 protocols using ab70895
1
Co-immunoprecipitation of USP10 and KLF4
2
Co-immunoprecipitation of USP10 and KLF4
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The interaction between USP10 and KLF4 was detected by Co-IP. BMSCs were lysed using Co-IP lysate to extract proteins. Protein supernatants were incubated with Normal rabbit IgG or USP10 (AB70895, Abcam), and SKP2 (AB214666, Abcam) antibodies at 4℃ overnight with rotation. The Co-IP lysate was then added to Protein A/G agarose beads, mixed, and centrifuged to collect the precipitate. Following antibody incubation, the cell lysate was fully incubated with agarose beads for 2 h at 4℃ to allow antibody coupling to the beads. The agarose beads were washed using Co-IP lysate buffer, and WB subsequently analyzed the coupling product.
3
Antibody Staining Protocol for Cellular Stress Granules
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The following antibodies were used: Mouse anti-eIF2a (1:1000 Cell Signaling L57A5), Mouse anti-G3BP (1 mg/ml BD Biosciences 611126), Rabbit anti-G3BP2 (1:1000 Bethyl A302-040A) Rabbit anti-PCNA (1 mg/ml Abcam ab18197), Rabbit anti-pS51-eIF2a (0.721 mg/ml ab32157), Mouse anti-Puromycin (1:25000 Sigma-Aldrich MABE343 [12D10]), Rabbit anti-TIA1 (5 mg/ml for staining and 1 mg/ml for western-blot Abcam ab40693), Mouse anti-aTubulin (1:1000 Sigma-Aldrich T9026), Rabbit anti-UBAP2L (0.5 mg/ml for staining and 1 ug/ml for western-blot Abcam ab138309), Rabbit anti-USP10 (5mg/ml Abcam ab70895), AlexaFluor donkey anti-mouse 488nm (4 mg/ml Thermo Fisher A21202), AlexaFluor donkey anti-rabbit 488nm (2 mg/ml Thermo Fisher A11034), AlexaFluor goat anti-rabbit 594nm (1 mg/ml Thermo Fisher A11012), AlexaFluor goat anti-rabbit 647nm (2 mg/ml Thermo Fisher A32733) HRP-conjugated goat anti-mouse (1:10000 BioRad 170-6516), HRP-conjugated goat anti-rabbit (1:10000 BioRad 170-6515).
For U-ExM both the primary and the secondary antibody concentrations were doubled. For STED the following secondary antibodies were used: Goat anti-mouse STAR RED (1:5000 Abberior STRED), Goat anti-mouse STAR RED (1:5000 Abberior STRED), AlexaFluor donkey anti-rabbit 488nm (1:5000 -0.2 mg/ml Thermo Fisher A11034).
For U-ExM both the primary and the secondary antibody concentrations were doubled. For STED the following secondary antibodies were used: Goat anti-mouse STAR RED (1:5000 Abberior STRED), Goat anti-mouse STAR RED (1:5000 Abberior STRED), AlexaFluor donkey anti-rabbit 488nm (1:5000 -0.2 mg/ml Thermo Fisher A11034).
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