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A770041

Manufactured by Axon Medchem
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Sourced in Netherlands

A770041 is a lab equipment product designed for use in scientific research and experiments. It serves as a core function in laboratory settings, but a detailed description without interpretation or extrapolation is not available.

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Lab products found in correlation

Human igd, by Abcam (2 mentions) P505 15, by Selleck Chemicals (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Streptavidin apc cy7, by BD (1 mentions) Human cd4 microbeads, by Miltenyi Biotec (1 mentions) Anti cd69 pe, by BD (1 mentions) Anti cd62l pe, by BD (1 mentions) Anti igd pe, by BD (1 mentions) Dasatinib, by Merck Group (1 mentions) Facs melody device, by BD (1 mentions) L glutamine, by Merck Group (1 mentions) Sodium pyruvate, by Merck Group (1 mentions) Il 17 pe, by BD (1 mentions) Ifn γ pe cy7, by BD (1 mentions) Il 4 apc, by BD (1 mentions) Cd154 pe, by BD (1 mentions) Foxp3 pe, by BD (1 mentions) Cd4 fitc, by BD (1 mentions) Cd25 apc, by BD (1 mentions)

6 protocols using a770041

1

Activation of Human T Cells by IgD

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Human IgD was purchased from Abcam (Cambridge, MA, United States). CP-25 was provided by the Chemistry Lab of the Institute of Clinical Pharmacology of Anhui Medical University with a purity of 98.8% (Hefei, China). CP-25 was dissolved in DMEM. Herbimycin A (HA) was purchased from Millipore (Temecula, CA, United States). A770041 was purchased from Axon Medchem (Groningen, Netherlands). Biotinylated IgD was prepared in our laboratory using a protein biotinylation kit from Pierce Biotechnology (Rockford, IL, United States) according to the manufacturer’s instructions. Human CD4 microbeads were purchased from Miltenyi Biotec (Germany). PE-anti-CD69, PE-anti-CD154, PE-anti-CD62L, PE-anti-IgD, PE-cy5-anti-CD4 monoclonal antibodies (mAbs), APC-Cy7-streptavidin and isotype-matched PE-labeled mouse IgG2a mAbs were purchased from BD Pharmingen (San Diego, CA, United States). The anti-Lck antibody was purchased from Cell Signaling Technology (Danvers, MA, United States).
Wu Y.J., Chen H.S., Chen W.S., Dong J., Dong X.J., Dai X., Huang Q, & Wei W. (2018). CP-25 Attenuates the Activation of CD4+ T Cells Stimulated with Immunoglobulin D in Human. Frontiers in Pharmacology, 9, 4.
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2

Establishment and Characterization of Human Head and Neck Cancer Cell Subclones

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The human squamous cell carcinoma cell lines FaDu (hypopharynx), Cal33 (oral tongue), XF354 (oral cavity) and SAS (oral tongue) were a kind gift of Prof. Daniel Zips and have been cultured in RPMI-1640 medium supplemented with 10% fetal bolvine serum (FBS), 100 Units/ml penicillin, 100 μg/ml streptomycin (Thermo Fisher Scientific, Waltham), 2 mM L-Glutamine and 1 mM sodium pyruvate (Sigma Life Science, St. Louis) at 37 °C and 5% CO2. SAS subclones were randomly isolated (no selection/surface marker) via single cell sorting into 96 well plates filled with culture medium containing 2.5 μg/ml amphotericin B using a FACS Melody device (BD Bioscience, New Jersey). A total of twenty subclones were further cultivated and used in subsequent experiments. For chemical LCK inhibition, cells were treated with dasatinib (Sigma-Aldrich, St. Louis) or A-770041 (Axon Medchem, Reston) as indicated.
Weiße J., Rosemann J., Müller L., Kappler M., Eckert A.W., Glaß M., Misiak D., Hüttelmaier S., Ballhausen W.G., Hatzfeld M., Haemmerle M, & Gutschner T. (2021). Identification of lymphocyte cell-specific protein-tyrosine kinase (LCK) as a driver for invasion and migration of oral cancer by tumor heterogeneity exploitation. Molecular Cancer, 20, 88.
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3

Immunological Analysis of T Cell Activation

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Human IgD was purchased from Abcam (Cambridge, MA, USA). Anti-CD3 and CD28 antibodies were provided by T&L Biological Technology (Beijing, China). A770041 was purchased from Axon Medchem (Groningen, Netherlands). rhTNFR:Fc fusion protein was purchased from Guojian Pharmaceutical Company (Shanghai, China). Anti-mouse IgD antibodies were purchased from eBioscience (San Diego, CA, USA). Anti-human CD69-PE-cy5, CD154-PE, CD4-FITC, CD25-APC, IL-4-APC, IFN-γ-PE-cy7, IL-17-PE, and FoxP3-PE, along with anti-mouse CD3e-PerCP-CyTM5.5, CD4-FITC, CD25-APC, CD154-PE, IFN-γ-PerCP-CyTM5.5, IL4-APC, IL-17-PE, and FoxP3-PE antibodies were provided by BD Pharmingen (San Diego, CA, USA). Anti-Lck, anti-phospho-ZAP70, anti-phospho-Lck, anti-ZAP70, and anti-β-actin were purchased from Affinity Biosciences, Sigma and Abcam.
Zhang J., Hu X., Dong X., Chen W., Zhang L., Chang Y., Wu Y, & Wei W. (2020). Regulation of T Cell Activities in Rheumatoid Arthritis by the Novel Fusion Protein IgD-Fc-Ig. Frontiers in Immunology, 11, 755.
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4

Phospho-protein Analysis of Leukemia Cells

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Murine and human leukemia cells pre-incubated in DMEM high glucose medium (Thermo Scientific) containing 10 % FBS for 1 hour at 37 °C, then treated for 30 minutes at 37°C using following small molecule inhibitors: dasatinib (LC laboratories, Woburn, MA), saracatinib (Selleckchem, Houston,TX), bosutinib (Selleckchem), p505-15 (Selleckchem), R406 (Selleckchem), R778 (fostamatinib, Selleckchem), RK24466 (Cayman Chemical, Ann Harbor, MI), LCKi-II (Fisher Scientific, Waltham, MA), A770041 (Axon Medchem, Reston,VA), ibrunitinib (PCI-32765, Selleckchem), buparlisib (Selleckchem), or trametinib (LC laboratories).
For intracellular staining, cells were fixed with 1.5% formaldehyde for 10 minutes at room temperature, permeabilized with 100% ice-cold methanol for 20 minutes on ice, and washed twice with staining buffer as described previously (9 (link),10 (link)), and stained with conjugated antibodies to intracellular phospho-proteins (Supplementary Table S2). Data were analyzed using Flowjo software.
Duque-Afonso J., Lin C.H., Han K., Wei M.C., Feng J., Kurzer J., Schneidawind C., Wong S.H., Bassik M.C, & Cleary M.L. (2016). E2A-PBX1 remodels oncogenic signaling networks in B-cell precursor acute lymphoid leukemia. Cancer research, 76(23), 6937-6949.
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5

Kinase Inhibitor Compound Characterization

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The 254 PKIs that were tested in this study constitute part of the Chemical Validation Library (CVL), which is the core library of the Nested Chemical Library of Vichem and consists of launched kinase inhibitor drugs and compounds in clinical trials and in preclinical development (http://www.vichem.hu/nested_chemical_library.html) [21] (link). Four of these 254 PKIs were further studied; PP2 is an inhibitor of Src family tyrosine kinases with higher potency for Lck and Fyn. Compound 5 also inhibits Src family kinases with Lck being the most sensitive. CI-1040 and PD 198306 are MEK inhibitors. PP2, CI-1040 and PD 198306 have already been used in pharmacological studies [22] (link)–[24] (link). Inhibitors were synthesized according to literature procedures: PP2 [25] (link), compound 5 [26] (link), CI-1040 and PD 198306 [27] (link). A-770041 (Axon Medchem) is a selective inhibitor of Lck, while U0126 (Gibco) is a selective inhibitor of MEK1 and MEK2. All inhibitors were dissolved in DMSO.
Mavromatidis V., Varga Z., Waczek F., Őrfi Z., Őrfi L., Kéri G, & Mosialos G. (2014). Identification of Protein Kinase Inhibitors with a Selective Negative Effect on the Viability of Epstein-Barr Virus Infected B Cell Lines. PLoS ONE, 9(4), e95688.
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6

Evaluating Targeted Therapies for Leukemia

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Leukemia cells were transduced with lentiviral vectors containing shRNA for luciferase (control) or indicated genes and mCherry fluorescence reporter. mCherry+ cells were sorted 7 days after transduction. Secondary bone marrow transplantation assay using 1000 mouse preBCR+/E2A-PBX1+ leukemia cells per recipient was described elsewhere (5 (link)). For in vivo treatment, mice were treated daily, intra-peritoneally, for 21 days, starting the day after transplantation with vehicle (30% PEG1500, 1% Tween 80, 2.5 % DMSO dissolved in PBS), 10 mg/kg b.w. p505-15 (Selleckchem) (11 (link)) or 5 mg/kg b.w. A770041 (Axon Medchem) (12 (link)).
Duque-Afonso J., Lin C.H., Han K., Wei M.C., Feng J., Kurzer J., Schneidawind C., Wong S.H., Bassik M.C, & Cleary M.L. (2016). E2A-PBX1 remodels oncogenic signaling networks in B-cell precursor acute lymphoid leukemia. Cancer research, 76(23), 6937-6949.
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