Recombinant Human MCL1, His-tagged

• Cat.No.: MCL1-779H
• Product Overview: Recombinant Human MCL1 protein, fused to His-tag, was expressed in E.coli and purified by Ni-sepharose.

Specification

• Source: E.coli
• Species: Human
• Tag: His
• Protein length: 1-350aa
• Storage: The protein is stored in PBS buffer at -20℃. Avoid repeated freezing and thawing cycles.
• Storage Buffer: 1M PBS (58mM Na2HPO4,17mM NaH2PO4, 68mM NaCl, pH8. ) added with 300mM Imidazole and 0.7% Sarcosyl, 15%glycerol.

Gene Information

• Gene Name: MCL1 myeloid cell leukemia sequence 1 (BCL2-related) [ Homo sapiens ]
• Official Symbol: MCL1
• Synonyms: MCL1; myeloid cell leukemia sequence 1 (BCL2-related); induced myeloid leukemia cell differentiation protein Mcl-1; BCL2L3; Mcl 1; bcl-2-like protein 3; myeloid cell leukemia ES; bcl-2-related protein EAT/mcl1; TM; EAT; MCL1L; MCL1S; Mcl-1; MCL1-ES; bcl2-L-3; mcl1/EAT; MGC1839; MGC104264
• Gene ID: 4170
• mRNA Refseq: NM_001197320
• Protein Refseq: NP_001184249
• MIM: 159552
• UniProt ID: Q07820
• Chromosome Location: 1q21
• Pathway: Apoptosis, organism-specific biosystem; Direct p53 effectors, organism-specific biosystem; E2F transcription factor network, organism-specific biosystem; HIF-1-alpha transcription factor network, organism-specific biosystem; IL-7 Signaling Pathway, organism-specific biosystem; IL6-mediated signaling events, organism-specific biosystem
• Function: BH3 domain binding; protein binding; protein channel activity; protein heterodimerization activity

Citation

Mcl-1 interacts with Akt to promote lung cancer progression

Journal: Cancer research    PubMed ID: 31662324    Data: 2020/6/15

Authors: Guo Chen, Dongkyoo Park, Xingming Deng

Article Snippet:BrdU cell proliferation kit and purified recombinant His tag-Akt protein (#14–276) were purchased from EMD Millipore (Billerica, MA).BrdU cell proliferation kit and purified recombinant His tag-Akt protein (#14–276) were purchased from EMD Millipore (Billerica, MA).. Purified recombinant His-tag-Mcl-1 protein (MCL1–781H) was obtained from Creative BioMart (Shirley, NY).. Mcl-1 rabbit polyclonal Ab (sc-819), anti-β-actin (sc-47778), anti-Bax (sc-493), anti-HA (sc-7392) and anti-GFP (sc-9996) antibodies as well as Akt1/2 siRNA (sc-43609) were purchased from Santa Cruz Biotechnology (Dallas, Texas).Mcl-1 rabbit polyclonal Ab (sc-819), anti-β-actin (sc-47778), anti-Bax (sc-493), anti-HA (sc-7392) and anti-GFP (sc-9996) antibodies as well as Akt1/2 siRNA (sc-43609) were purchased from Santa Cruz Biotechnology (Dallas, Texas).

Knockout of Mcl-1 suppresses cancer cell growth via downregulation of Akt activity. A-E, Mcl-1 was knocked out from H1299 cells using CRISPR/Cas9, followed by analysis of Mcl-1 expression by Western blot (A), growth curve (B), colony formation (C), human phospho-kinase array (1: Akt S473; 2: Akt T308) (D), and quantification of human phospho-kinase by image J (E). Data represent the mean ± SD. *P < 0.05, ***P < 0.001, by 2-tailed t test. (F) Mcl-1 was knocked out from H1299 cells, or knocked down from H460 cells, followed by Western blotting analysis of Mcl-1, pAkt, pmTOR and p-p70S6K.

The binding of Mcl-1 with Akt is essential for Mcl-1 to disrupt PH/KD interactions, upregulate Akt activity and promote tumor growth. A and B, GST-tagged kinase domain of Akt and GFP-tagged PH domain of Akt were co-transfected with Flag-tagged WT Mcl-1 or ΔPEST Mcl-1 mutant into H1299 cells, followed by GST pull down (A), or co-IP using GFP antibody (B), and Western blot using GST or GFP antibody. C, Interaction between the kinase domain and PH domain in the absence or presence of WT Mcl-1 or ΔPEST was measured in a mammalian two-hybrid system. D and E, H1299 Mcl-1?/? cells were transfected with empty vector (EV), Flag-tagged WT Mcl-1 or ΔPEST Mcl-1 mutant, followed by analysis of pAkt, pmTOR and p-p70 S6K by Western blot (D) and colony formation assay (E). Data represent the mean ± SD. ***P < 0.001, by 2-tailed t test. F, The same number (3×106) of H1299 Mcl-1?/? cells expressing EV, Flag-tagged WT Mcl-1 or ΔPEST mutant were injected into subcutaneous tissue in the flank region of nude mice to generate lung cancer xenografts. Tumor volume was measured once every 3 days. After 33 days, the mice were sacrificed and the tumors were removed, photographed, and analyzed. Data represent the mean ± SD, n=6 mice per group. ***P < 0.001, by 2-tailed t test.

Knockout of Mcl-1 inhibits tumor growth in vivo. A and B, The same number (3×106) of H1299 parental or Mcl-1?/? cells were injected into subcutaneous tissue in the flank region of nude mice to generate lung cancer xenografts (n=5 mice each group). Tumor volume was measured once every 3 days. After 33 days, the mice were sacrificed and the tumors were removed, photographed, and analyzed. Data represent the mean ± SD, n=5 per group. ***P < 0.001, by 2-tailed t test. C-H, IHC staining of Mcl-1, pAkt (S473), pAkt (T308), Ki67 and active caspase 3 was performed in tumor tissues at the end of experiments. Data represent the mean ± SD, n=5 per group. **P < 0.01, ***P < 0.001, by 2-tailed t test.

First evidence that oligopyridines, α-helix foldamers, inhibit Mcl-1 and sensitize ovarian carcinoma cells to Bcl-xL-targeting strategies.

Journal: Journal of medicinal chemistry    PubMed ID: 25585174    Data: 2015/5/7

Authors: Céline Gloaguen, Anne Sophie Voisin-Chiret, Laurent Poulain

Article Snippet:Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.. Recombinant human Mcl-1 protein, fused to His-tag, and recombinant human Bcl-xL, His-tagged (DNA sequence encoding the human BCL2L1 isoform 1 (Met 1?Arg 212) fused with a polyhistidine tag at the C-terminus) were purchased from Creative BioMart (45?16 Ramsey Road, Shirley, NY 11967, USA).. A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.

Reviews

06/20/2022

Another advantage of using MCL1 protein in trials is its potential as a therapeutic agent.

01/16/2022

The manufacturer's support and expertise further enhance the utility of MCL1 in my experimental work, ensuring reliable and high-quality results.

03/23/2016

This collaborative partnership greatly contributes to the success of my research projects and facilitates meaningful contributions to the scientific community.

Q&As

Can targeting MCL1 have implications beyond cancer therapy?

Yes, MCL1 modulation may have therapeutic implications in other diseases, such as neurodegenerative disorders, where apoptosis is a contributing factor.

Are there any diagnostic tools or biomarkers that can assess MCL1 expression in cancer patients?

Immunohistochemistry and molecular assays can be used to measure MCL1 expression levels, providing insights into the prognosis and potential treatment strategies.

How does the understanding of MCL1 function contribute to personalized medicine in cancer treatment?

Understanding MCL1 expression allows for the identification of patients who may benefit from targeted therapies, contributing to personalized treatment strategies.

Are there any ongoing clinical trials exploring MCL1 inhibitors as a therapeutic approach?

Yes, there are several clinical trials investigating the use of MCL1 inhibitors in different cancer types to assess their safety and efficacy.

How does the MCL1 protein interact with other proteins in the Bcl-2 family?

MCL1 interacts with pro-apoptotic and anti-apoptotic proteins in the Bcl-2 family, influencing the balance between cell survival and death.