Anti rack1

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-RACK1 is a primary antibody that detects the receptor for activated C kinase 1 (RACK1) protein. RACK1 is a member of the tryptophan-aspartate repeat (WD repeat) protein family and functions as a scaffolding protein, facilitating the interaction of various signaling molecules. This antibody can be used in applications such as western blotting to identify and quantify RACK1 expression levels in biological samples.

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3 protocols using anti rack1

Comprehensive Western Blotting Protocol

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Western blotting was performed as described previously (Johnson et al. 2018 (link)). When reprobing blots, HRP-conjugated secondary antibodies were either inactivated by incubation with sodium azide in 5% skim milk/TBST or stripped with Restore Stripping Buffer (ThermoFisher). The following antibodies were used in this study: anti-RACK1 (Cell Signaling, ref. 4716S, 1:1000), anti-RACK1 (Santa Cruz, ref. 17754, 1:500), RPL5 (Genetex, ref. 101821, 1:1000), uS10/RPS20 (abcam, ref. 133776, 1:1000), eS10/RPS10 (Genetex, ref. 101836, 1:1000), uS5/RPS2 (Santa Cruz, ref. 130399, 1:500), and uS3/RPS3 (Bethyl, ref. 303-840A, 1:1000).

Johnson A.G., Lapointe C.P., Wang J., Corsepius N.C., Choi J., Fuchs G, & Puglisi J.D. (2019). RACK1 on and off the ribosome. RNA, 25(7), 881-895.

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Immunohistochemical Analysis of Protein Expression

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Briefly, formalin-fixed, paraffin-embedded tissue sections were dewaxed with xylene. After rehydration and antigen retrieval, sections were blocked with 5% BSA for 60 min and then incubated with the primary antibodies at 4 °C overnight. The antibodies included: anti-TRIM26 antibody (1:200, Santa Cruz Biotechnology), anti-RACK1 (1:200, Cell Signaling Technology), anti-p-MEK1/2 (1:200, Cell Signaling Technology), anti-p-ERK1/2 (1:200, Cell Signaling Technology), anti-E-cadherin (1:200, Cell Signaling Technology), anti-N-cadherin (1:200, Cell Signaling Technology), and anti-vimentin (1:200, Cell Signaling Technology). The next day, sections were stained with diaminobenzidine followed by counterstaining with hematoxylin and washing in water. The sections were observed and photographed under a ×200 objective by light microscopy (Olympus, Japan).

Xia K., Zheng D., Wei Z., Liu W, & Guo W. (2023). TRIM26 inhibited osteosarcoma progression through destabilizing RACK1 and thus inactivation of MEK/ERK signaling. Cell Death & Disease, 14(8), 529.

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Protein Expression and Detection

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Preparation of whole cell lysates and western blot analysis was performed as described. The primary antibodies used in this study were anti-EGFP (1:1000, Origene, USA), anti-HA (1:1000, Origene, USA), anti-Flag (1:1000, Santa Cruz, USA), anti-LC3 (1:1000, Cell signaling, USA), anti-RACK1 (1:1000, Cell signaling, USA) and anti-Actin (1:1000, Santa Cruz, USA).

An N., Li B., Liu Y., Zhang Z., Su R., & Liu X. (2021). The Surface Antigen 1(SAG1) of Toxoplasma Gondii Interacts With RACK1 Inducing Autophagy and Maintaining The Viability of Host Cells.

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