Anti ck2α

Manufactured by Cell Signaling Technology

Sourced in Japan, China

Anti-CK2α is a primary antibody that specifically recognizes the CK2α subunit of the protein kinase CK2. CK2α is a highly conserved serine/threonine protein kinase that plays a crucial role in various cellular processes.

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Lab products found in correlation

4 protocols using anti ck2α

Immunoblotting and Immunoprecipitation of Cortactin and AKT

Cited 2 times

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Western blotting was conducted as described (27 (link)) and visualized with autoradiography film (E3012, Denville Scientific) or captured by an Amersham Imager 600 (GE Healthcare Bio-Sciences). Antibodies used were: anti-cortactin clone 4F11 (1 μg/ml, (26 (link))), anti-pS473 AKT (#4060, 1:1000; Cell Signaling Technology), anti-panAKT (#2920, 1:1000; Cell Signaling Technology), anti-β-actin (#8457, 1:1000; Cell Signaling Technology), anti-CK2α (#2656, 1:500, Cell Signaling Technology), anti-DYKDDDK (FLAG) clone 2EL-1B11 (MAB3118, 1:500, Millipore) and anti-Arp3 (#07-272, 1:500, EMD Millipore). Immunoprecipitation was conducted from cells lysed in 50 mM Tris Buffer pH 8.0 with 10 mM EDTA and 1% NP-40 (28 (link)). Clarified lysates (1 mg) were incubated with 50 μl of FLAG M2 affinity resin (A2220, Sigma-Aldrich) for 2 hours at 4°C. Immune complexes were collected by centrifugation, washed twice with Tris buffer, separated by SDS-PAGE, and Western blotted with antibodies as described above.

Markwell S.M., Ammer A.G., Interval E.T., Allen J.L., Papenberg B.W., Hames R.A., Castaño J.E., Schafer D.A, & Weed S.A. (2019). Cortactin Phosphorylation by Casein Kinase 2 Regulates Actin-Related Protein 2/3 Complex Activity, Invadopodia Function and Tumor Cell Invasion. Molecular cancer research : MCR, 17(4), 987-1001.

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Western Blot Analysis of Autophagy Signaling

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Cells were lysed in RIPA buffer (Thermo Scientific, Rockford, IL) with the cocktail of protease and phosphatase inhibitors PhoSTOP (Roche, Mannheim, Germany). Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, transferred to NitroPure membranes (Macherey-Nagel, Dϋren, Germany) and blocked with 5% BSA (Thermo Scientific, Rockford, IL) in PBS/0.1% Tween. Blots were incubated with primary antibodies (Cell Signaling Technology, Beverly, MA): rabbit polyclonal anti-PARP (1:1000), rabbit polyclonal anti-CK2α (1:1000), rabbit polyclonal anti-LC3 (1:1000), rabbit monoclonal anti-pSer757-ULK-1 (1:1000), rabbit monoclonal anti-ULK-1 (1:1000), rabbit monoclonal anti-Akt (1:1000), rabbit monoclonal anti-pThr389-S6K1 (1:1000), rabbit polyclonal anti-S6K1 (1:1000), rabbit polyclonal anti-pSer235/Ser236-S6 (1:1000) and mouse monoclonal anti-S6 (1:1000). Mouse monoclonal anti-p62 (1:1000) was obtained from BD Biosciences and anti-actin (1:2000) from Santa Cruz Biotechnology (Santa Cruz, CA). The secondary antibodies used were: anti-goat IgG-HRP (1:2000), anti-rabbit IgG-HRP (1:2000) and anti-mouse IgG-HRP (1:2000) from Santa Cruz Biotechnology. Blots were revealed with the EZ-ECL chemiluminescent kit (Biological Industries, Haemek, Israel).

Silva-Pavez E., Villar P., Trigo C., Caamaño E., Niechi I., Pérez P., Muñoz J.P., Aguayo F., Burzio V.A., Varas-Godoy M., Castro A.F., Colombo M.I, & Tapia J.C. (2019). CK2 inhibition with silmitasertib promotes methuosis-like cell death associated to catastrophic massive vacuolization of colorectal cancer cells. Cell Death & Disease, 10(2), 73.

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Antibody Validation for Molecular Analyses

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The following antibodies (Abs) were used for western blotting, immunoprecipitation, qPCR, ELISA, confocal microscopy, immunohistochemistry, flow cytometry, or joint injection: anti-TMEM147 (sc-138814, Santa Cruz Biotechnology), anti-TMEM147 (generated by immunizing rabbit with peptide containing amino acids 121-136 VGARGIEFDWKYIQMSC, Sigma-Aldrich), anti-NF-κB p65 (sc-372, Santa Cruz Biotechnology), anti-phospho-p65 S536 (#3033, Cell Signaling Technology, Tokyo, Japan), anti-phospho NF-κB p65 S529 (ab47395, Abcam, Tokyo, Japan), anti-phospho NF-κB p65 S276 (SAB4504488, Sigma-Aldrich), anti-phospho-stat3 (#9145, Cell Signaling Technology), anti-IκBα (#4814, Cell Signaling Technology), anti-phospho-IκBα (#9246, Cell Signaling Technology), anti-BCR (#3902, Cell Signaling Technology), anti-phospho-BCR Y177 (#3901, Cell Signaling Technology), anti-CK2α (#2656, Cell Signaling Technology), anti-phospho-CK2α (SAB4300628, Sigma-Aldrich), anti-α-tubulin (T5168, Sigma-Aldrich), anti-FLAG M2 (F1804, Sigma-Aldrich), rabbit IgG (#3900, Cell Signaling Technology), anti-rabbit IgG (sc-2004, Santa Cruz Biotechnology), anti-mouse IgG (sc-2314, Santa Cruz Biotechnology), anti-goat IgG (sc-3851, Santa Cruz Biotechnology), Alexa Fluor 488 goat anti-rabbit IgG (H+L) (A11055, Life technologies, Tokyo, Japan) and Hoechst 33342 trihydrochloride trihydrate (H3570, Life technologies).

Ota M., Tanaka Y., Nakagawa I., Jiang J.J., Arima Y., Kamimura D., Onodera T., Iwasaki N, & Murakami M. (2020). Role of Chondrocytes in the Development of Rheumatoid Arthritis Via Transmembrane Protein 147-Mediated NF-κB Activation. Arthritis & rheumatology (Hoboken, N.J.), 72(6).

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Western Blot Analysis of DNA Repair Proteins

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Whole cell lysates were prepared with RIPA lysis buffer containing a protease inhibitor cocktail (Roche, Manheim, Germany). Western blotting analysis was performed as previously described 15 (link). The following antibodies were used as follows: monoclonal anti-JWA (1:500, contract produced by AbMax, Beijing, China); polyclonal anti-PARP-1, anti-γ-H2AX, anti-XRCC1, anti-Bcl-2, anti-CK2α, anti-CAT, anti-Nrf2, anti-p-XRCC1 and p-ERK (1:1000-2000, Cell Signaling Technology, Danvers, MA, USA). Tubulin and GAPDH were used as loading controls.

Zhang Y., Chen J., Che Z., Shu C., Chen D., Ding K., Li A, & Zhou J. (2021). JP3 enhances the toxicity of cisplatin on drug-resistant gastric cancer cells while reducing the damage to normal cells. Journal of Cancer, 12(7), 1894-1906.

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