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Anti rack1

Manufactured by Santa Cruz Biotechnology
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Anti-RACK1 is a laboratory reagent that acts as an antibody specific to the RACK1 (Receptor for Activated C Kinase 1) protein. RACK1 is a highly conserved intracellular protein that serves as a scaffolding molecule, facilitating the interactions between various signaling proteins. The Anti-RACK1 antibody can be used to detect and study the expression, localization, and interactions of the RACK1 protein in biological samples.

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Lab products found in correlation

Anti cbp, by Abcam (1 mentions) Anti xpress, by Thermo Fisher Scientific (1 mentions) Anti gemin5, by Novus Biologicals (1 mentions) Anti gst antibody, by Santa Cruz Biotechnology (1 mentions) Anti tubulin, by Merck Group (1 mentions) Anti rack1, by Cell Signaling Technology (1 mentions) Restore stripping buffer, by Thermo Fisher Scientific (1 mentions) Anti cpne1, by Abcam (1 mentions) Anti pkr, by Santa Cruz Biotechnology (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti phospho eif2α ser51, by Cell Signaling Technology (1 mentions) Anti ha hrp, by Roche (1 mentions) Z vad fmk, by Santa Cruz Biotechnology (1 mentions) Anti pkr, by Abcam (1 mentions) Anti g3bp1 antibody, by BD (1 mentions) Anti actin, by Cell Signaling Technology (1 mentions) Anti parp, by Cell Signaling Technology (1 mentions) Isrib, by Merck Group (1 mentions) 3 methyladenine, by Merck Group (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions)

7 protocols using anti rack1

1

Immunodetection of G5 Proteins

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The immunodetection of G5845-1508 proteins were performed with different antibodies depending on the protein tag (anti-CBP [Abcam] for G5845-1508-TAP, anti-Xpress [Thermo Fisher Scientific] for Xpress-G5845–1508, or anti-His [Merck] for His-G5845–1508). The anti-Gemin5 (Novus) antibody was used to detect the endogenous Gemin5 protein. The endogenous proteins tubulin, RACK1, and P0 were immunodetected with anti-Tubulin (Merck), anti-RACK1 (Santa Cruz), and 3BH5 (anti-P0) (Vilella et al, 1991 (link)) antibodies. GST fusion proteins were detected with the anti-GST antibody (Santa Cruz).
Francisco-Velilla R., Embarc-Buh A., del Caño-Ochoa F., Abellan S., Vilar M., Alvarez S., Fernandez-Jaen A., Kour S., Rajan D.S., Pandey U.B., Ramón-Maiques S, & Martinez-Salas E. (2022). Functional and structural deficiencies of Gemin5 variants associated with neurological disorders. Life Science Alliance, 5(7), e202201403.
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2

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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3

Immunocytochemistry of CPNE1 and RACK1

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Cells were seeded in 12-well plates containing pre-inserted glass slides. Then, the cells were washed with PBS 24 h later at a confluence of 40–50%. Cells were fixed with 4% paraformaldehyde for 30 min afterwards, followed by permeabilization with 0.5% Triton X-100 solution for an additional 20 min. Next, 5% bovine serum albumin was added to function as the blocking buffer. The primary antibodies, anti-CPNE1 (Abcam, UK) and anti-RACK1 (Santa Cruz, USA), and corresponding secondary antibodies conjugated to Cy3 and FITC were used successively.
Wang A., Yang W., Li Y., Zhang Y., Zhou J., Zhang R., Zhang W., Zhu J., Zeng Y., Liu Z, & Huang J.A. (2022). CPNE1 promotes non-small cell lung cancer progression by interacting with RACK1 via the MET signaling pathway. Cell Communication and Signaling : CCS, 20, 16.
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4

Antibodies and Reagents in Cell Study

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Antibodies and reagents used in this study were as follows: anti-G3BP1 antibody (BD Bioscience, 611126, Franklin Lakes, NJ, USA), anti-PKR (Santa cruz biotechnology, sc-6282, Dallas, TX, USA) anti-RACK1 (Santa Cruz biotechnology sc-17754), anti-phospho-eIF2α (Ser51) (Cell Signaling Technology, 9721S, Danvers, MA, USA), anti-cleaved caspase-3 (Cell Signaling Technology, 9661S), anti-PARP (Cell Signaling Technology, 9542S), anti-Actin (Cell Signaling Technology, 3700S), anti-PKR (Abcam, ab52506, Cambridge, MA, USA), anti-phospho-PKR(Thr451) (Abcam, ab81303), anti-IMP1 antibody (Bethel Laboratories, A303-424A, Montgomery, TX, USA), anti-HA-HRP (Roche, 12013819001, Mannheim, Germany), morusin (root bark of Morus alba) (Biopurity Phytochemicals Ltd., BP0961, Chengdu, Sichuan, China), ISRIB (Millipore sigma, SML0843, Saint Louis, MO, USA), 3-methyladenine (Millipore sigma, M9281), zVAD-FMK (Santa cruz biotechnology, sc-3067).
Park Y.J., Choi D.W., Cho S.W., Han J., Yang S, & Choi C.Y. (2020). Stress Granule Formation Attenuates RACK1-Mediated Apoptotic Cell Death Induced by Morusin. International Journal of Molecular Sciences, 21(15), 5360.
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5

Western Blot Protocol for Protein Expression

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Protein extracts of different samples including tissues and cells were first quantified using a BCA protein assay kit, mixed with Laemmli's loading buffer, applied to 10% SDS‐PAGE, and then transferred onto a PVDF membrane. The blots were probed with the anti‐RACK1, anti‐GAPDH, anti‐β‐actin anti‐cyclin D1, purchased from Santa Cruz Biotechnology, anti‐p21, anti‐p27, anti‐p‐Akt and anti‐Akt antibodies purchased from Cell Signaling Technology, followed by the horseradish (HRP)‐conjugated secondary antibody (1:5000); finally, the immunoreactions were visualized by ECL and exposed to X‐ray film.
Yang D., Lu X., Zhang X., Zhang X., Zhu L, & Liu Q. (2024). RACK1 promotes the occurrence and progression of cervical carcinoma. Journal of Clinical Laboratory Analysis, 38(4), e25012.
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6

Immunohistochemical Profiling of Brain Proteins

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AD and control human brain sections were deparaffinized in xylene and rehydrated through graded ethanol series. Antigen retrieval was performed by incubating the slides in a steamer with citrate buffer (pH 6.0) (Sigma) for 30 min. The slides were then washed for 10 min in 1 × PBS and incubated in 3% H2O2 (in methanol) for 10 min in order to block any endogenous peroxidase activity. Slides were then washed with 1 × PBS for 10 min. To prevent nonspecific binding and excessive background, slides were blocked with a serum-free buffer for 30 min. Primary antibodies (Anti-Rack1, Santa Cruz sc-17754, 1:500; Anti-ZNF598, GeneTex GRX119245, 1:250; Anti-ABCE1, Gift from Dr. Hegde; Anti-NEMF, ProteinTech 11,840-AP, 1:500; Anti-RPL22, ProteinTech 25,002-1-AP, 1:1000; Anti-ANKZF1, ProteinTech 20,447–1-AP, 1:1000) were applied on the slides and incubated overnight at 4 °C. Slides were washed with PBS and incubated with secondary antibody (conjugated to fluorophores) for 2 h at room temperature. After washing with 1 × PBS, mounting medium was used to mount the slides. Images were taken by Leica SP8 microscope.
Rimal S., Li Y., Vartak R., Geng J., Tantray I., Li S., Huh S., Vogel H., Glabe C., Grinberg L.T., Spina S., Seeley W.W., Guo S, & Lu B. (2021). Inefficient quality control of ribosome stalling during APP synthesis generates CAT-tailed species that precipitate hallmarks of Alzheimer’s disease. Acta Neuropathologica Communications, 9, 169.
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7

Antibody Validation for Protein Analysis

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Anti-GPS2 antibody was generated in rabbit against a peptide representing aa 307-327.
Commercial antibodies used were as follows: anti-ubiquitin (P4D1 clone, Cell Signaling Technology), anti-β-tubulin (TUB 2.1 clone, Sigma), anti-HDAC2 (ab16032, Abcam), anti-mtHSP70 (catalog no. MA3-028, Invitrogen), anti-K63 (catalog no. 05-1308, Millipore), anti-α-Tubulin (catalog no. T5168, Sigma), anti-GAPDH (catalog no. MA5-15738, Invitrogen), anti-PABPC1 (ab21060, Abcam), anti-RPS11(NBP2-22289, Novus Biologicals), anti-Rack1 (sc-17754, Santa Cruz Biotechnology), anti-Eif4G (15704-1-AP, Proteintech), anti-Paip1 (sc-365687, Santa Cruz Biotechnology), anti-Paip2 (15583-1-AP, proteintech), anti-Flag-HRP (catalog no. a8592, Sigma), anti-Puromycin (clone 12D10, Millipore), anti-PDHA (ab168379, Abcam), anti-Oxphos (ab110413, Abcam), anti-SOD2 (sc-137254, Santa Cruz Biotechnology). siRNAs against mouse GPS2, UBC13, MKRN1, MUL1, and UBC9 were purchased from Ambion. Nonspecific scrambled siRNA was included as negative controls in each experiment.
Gao Y., Cardamone M.D., Kwan J., Orofino J., Hekman R., Lyons S.M., Emili A., & Perissi V. (2021). Inhibition of Mul1-mediated ubiquitination promotes mitochondria-associated translation.
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