Limk 1

Manufactured by Santa Cruz Biotechnology

Sourced in United Kingdom

LIMK-1 is a protein kinase enzyme that plays a role in the regulation of actin cytoskeleton dynamics. It is involved in the phosphorylation and inactivation of cofilin, an actin-depolymerizing factor. The core function of LIMK-1 is to modulate the organization and stability of the actin cytoskeleton.

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Close spelling variants of this product

P-LIMK-1/2

3 protocols using limk 1

Luteolin and BMS-5 Cellular Assays

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Luteolin (purity ≥ 98%) was purchased from Sichuan Weikeqi Biological Technology CO., Ltd (CAS#: 491‐70‐3). BMS‐5 (purity ≥ 98%) was purchased from Enzo Life Sciences (CAS#:1338247‐35‐0). CNBr‐activated Sepharose^TM 4B (Lot# 10265330) was purchased from GE Healthcare. Thiazolyl blue tetrazolium bromide (MTT) powder was purchased from Solarbio Technology Co., Ltd. Dimethylsulfoxide (DMSO) was purchased from Sigma. Cyclin D1 (catalog# 2922), cyclin D3 (catalog# 2936), Bax (catalog# 5023), cleaved‐PARP (catalog# 5625), caspase‐3 (catalog# 9662), cleaved caspase‐3 (catalog# 9664), caspase‐7 (catalog# 9492), cleaved caspase‐7 (catalog# 8438), ROCK1 (catalog# 4035) and ROCK2 (catalog# 9029) were purchased from Cell Signaling Technology. Phospho‐LIMK‐1/2 (Thr 508/505)‐R (catalog# sc‐28409‐R), LIMK‐1(catalog# sc‐28370), cofilin (catalog# sc‐33779) and phospho‐cofilin (mSer3)‐R (catalog# sc‐21867‐R) were purchased from Santa Cruz Technology.

Zhang M., Wang R., Tian J., Song M., Zhao R., Liu K., Zhu F., Shim J., Dong Z, & Lee M. (2021). Targeting LIMK1 with luteolin inhibits the growth of lung cancer in vitro and in vivo. Journal of Cellular and Molecular Medicine, 25(12), 5560-5571.

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Cell Lysis and Western Blotting Procedure

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Cells were lysed in standard lysis buffer (SLB; 10 mM K₂HPO₄, 150 mM NaCl, 5 mM EDTA, 5 mM EGTA, 0.5% Triton X-100, 0.2% sodium deoxycholate; pH 7.4) supplemented with protease (cOmpleteTM, mini, EDTA-free; Roche, Mannheim, Germany; #1183617001) and phosphatase (PhosSTOP EASY pack; Roche, #04906837001) inhibitors, if not stated otherwise.
Western blotting was carried out as previously described [34 (link),36 (link),37 (link),83 (link),93 (link),94 (link)] using the following primary and secondary antibodies: Cofilin (1:2000, Cell Signaling, Frankfurt am Main, Germany; D3F9, #5175), p-Cofilin (Ser3, 1:1000, Cell Signaling, 77G2, #3313), Hsp60 (1:1000, Cell Signaling, D307, #4870), Pan-Cadherin (1:1000, Abcam, Cambridge, UK; ab6528), LIMK-1 (1:500, Santa Cruz Biotechnology, Heidelberg, Germany; H-84, sc-5576), p-LIMK-1/2 (Thr508/505, 1:200, Santa Cruz, sc-28409-R), POD-F(ab’)₂-anti-Rabbit IgG (H+L, 1:5000, Jackson Immuno Research, Ely, UK; #711-036-152), and POD-anti-Mouse IgG (H+L, 1:5000, Jackson Immuno Research, #715-035-151). Signals were detected using Immobilon^® Western Chemiluminescent HRP Substrate (Merck, Darmstadt, Germany; #WBKLS0500) and an Odyssey^® Fc detection system (Li-Cor Biosciences, Bad Homburg vor der Höhe, Germany). Image processing and densitometric analysis was performed with the Li-Cor software Image Studio Lite (Ver. 5.2.5).

Baltzer S., Bulatov T., Schmied C., Krämer A., Berger B.T., Oder A., Walker-Gray R., Kuschke C., Zühlke K., Eichhorst J., Lehmann M., Knapp S., Weston J., von Kries J.P., Süssmuth R.D, & Klussmann E. (2022). Aurora Kinase A Is Involved in Controlling the Localization of Aquaporin-2 in Renal Principal Cells. International Journal of Molecular Sciences, 23(2), 763.

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Analyzing Dasatinib-Induced Apoptosis

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Dasatinib (purity ≥ 99%) was purchased from Santa Cruz Biotechnology (CAS: 302962-49-8, Lot# H0515) (Beverly, MA, United States). BMS-5 (purity ≥ 98%) was bought from Enzo Life Sciences (CAS:1338247-35-0) (Shanghai, China). Dimethylsulfoxide (DMSO) and CNBr-activated Sepharose^TM 4B (Lot# 10265330) were purchased from Sigma (St. Louis, MO, United States) and GE Healthcare (Uppsala, Sweden) respectively. Cyclin D1 (Cat# 2922), cyclin D3 (Cat# 2936), cleaved-PARP (Cat# 5625), caspase-3 (Cat# 9662), cleaved caspase-3 (Cat# 9664) caspase-7 (Cat# 9492), and cleaved caspase-7 (Cat# 8438) were purchased from Cell Signaling Technology (Beverly, MA, United States). p-LIMK (Thr 508/505)-R (Catalog# sc-28409-R), LIMK-1 (Catalog# sc-28370), Cofilin (Catalog# sc-33779), p-Cofilin (mSer3)-R (Catalog# sc-21867-R) were purchased from Santa Cruz Technology. β-actin antibody (Cat# TA-09) as a loading control was obtained from ZSGB-Bio Company (Beijing, China).

Zhang M., Tian J., Wang R., Song M., Zhao R., Chen H., Liu K., Shim J.H., Zhu F., Dong Z, & Lee M.H. (2020). Dasatinib Inhibits Lung Cancer Cell Growth and Patient Derived Tumor Growth in Mice by Targeting LIMK1. Frontiers in Cell and Developmental Biology, 8, 556532.

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