Human ACK1 Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human ACK1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of ACK1 protein (Cat: 11080-H09B) from the overexpression lysate was verified.
A DNA sequence encoding the amino acid (Gly 110-Trp 476) of human ACK1 isoform 1 (NP_005772.3) was expressed with the GST tag at the N-terminus.
The recombinant human ACK1/GST chimera consists of 592 amino acids and predicts a molecular mass of 68 kDa. It migrates as an approximately 62 kDa band in SDS-PAGE under reducing conditions.
Human ACK1 Baculovirus-Insect cells Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human ACK1 Baculovirus-Insect cells Overexpression Lysate: Alternative Names
Human ACK Overexpression Lysate; Human ACK-1 Overexpression Lysate; Human ACK1 Overexpression Lysate; Human p21cdc42Hs Overexpression Lysate
ACK1 Background Information
ACK1 (also known as ACK, TNK2, or activated Cdc42 kinase) is a structurally unique non-receptor tyrosine kinase that is expressed in diverse cell types. This downstream effector of CDC42 which mediates CDC42-dependent cell migration via phosphorylation of BCAR1. The ACK1 protein may be involved in a regulatory mechanism that sustains the GTP-bound active form of Cdc42Hs and which is directly linked to a tyrosine phosphorylation signal transduction pathway. ACK1 integrates signals from plethora of ligand-activated receptor tyrosine kinases (RTKs), for example, MERTK, EGFR, HER2 and PDGFR to initiate intracellular signaling cascades. It binds to both poly- and mono-ubiquitin and regulates ligand-induced degradation of EGFR. ACK1 transduces extracellular signals to cytosolic and nuclear effectors such as the protein kinase AKT/PKB and androgen receptor (AR), to promote cell survival and growth. ACK1 participates in tumorigenesis, cell survival, and migration. Gene amplification and overexpression of ACK1 were found in many cancer types such as those of the lung and prostate. Recently, four somatic missense mutations of ACK1, which occur in the N-terminal region, the C-lobe of the kinase domain, and the SH3 domain, were identified in cancer tissue samples.
tyrosine kinase, non-receptor, 2
Mahajan K, et al. (2010) Shepherding AKT and androgen receptor by Ack1 tyrosine kinase. J Cell Physiol. 224(2): 327-33.
Chua BT, et al. (2010) Somatic mutation in the ACK1 ubiquitin association domain enhances oncogenic signaling through EGFR regulation in renal cancer derived cells. Mol Oncol. 4(4): 323-34.
Prieto-Echage V, et al. (2010) Cancer-associated mutations activate the nonreceptor tyrosine kinase Ack1. J Biol Chem. 285(14): 10605-15.
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