G7781

Manufactured by Merck Group

Sourced in United States

G7781 is a laboratory equipment product manufactured by Merck Group. It is designed for use in scientific research and analysis applications. The core function of G7781 is to perform precise measurements and data collection tasks in a controlled laboratory environment.

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Lab products found in correlation

8 protocols using g7781

Immunoblotting and Immunofluorescence Protocols

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Primary antibodies for immunoblotting: mouse anti-Ub (FK2; BML-PW8810 and UBCJ2; AB_2935893, Enzo Life Science), mouse anti-DnaK (8E2/2, ADI-SPA- 880, Enzo Life Science), goat anti-GroEL (ABIN6292975, antibodies-online), mouse anti-tubulin (E7, DSHB), rabbit anti-GST (G7781, Sigma) and rat anti-GFP (3H9, Proteintech).
Primary antibodies used for immunofluorescence microscopy: α-Ub (1:400, FK2; BML-PW8810 and UBCJ2; AB_2935893, Enzo Life Science) α-M1 (1:400, 1E3, ZRB2114, Merck) α-K63 (1:400, Apu3, 05–1308, Millipore) and α-LC3 (1:300, CTB-LC3–2-IC, Cosmo Bio,). Anti-Burkholderia antibody was provided by the United States Army Medical Research Institute of Infectious Diseases, an agency of the U.S. Government (“USAMRIID”).
Secondary antibodies used: Thermo Fisher Scientific (1:500, Alexa-conjugated anti-mouse, anti-goat and anti-rabbit antisera) and Dabco (1:5000, HRP-conjugated reagents).

Szczesna M., Huang Y., Lacoursiere R.E., Bonini F., Pol V., Koc F., Ward B., Geurink P.P., Pruneda J.N, & Thurston T.L. (2023). Dedicated bacterial esterases reverse lipopolysaccharide ubiquitylation to block immune sensing. Research Square.

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Immunofluorescence Assay for p65, FLAG, and GST

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MCF10A cells seeded on glass coverslips in 24-well plates (5.0 × 10⁴ cells per well) were transfected as described above. Twenty-four hours after transfection, or after TNFα addition when stated, cells were fixed for 10 min in 4% paraformaldehyde (PFA). Coverslips were then sequentially incubated with PBS-Triton X-100 for 5 min, and with antibodies against p65 (SantaCruz Biotechnology #sc-372), FLAG (Sigma-Aldrich #F1804) or GST (Sigma-Aldrich #G7781) diluted in PBS–BSA 3% for 1 h. Secondary antibodies anti-rabbit-AlexaFluor-555 (Cell Signaling #4413) and anti-mouse-AlexaFluor-488 (Cell Signaling #4408) were next incubated in PBS–BSA 3% for 1 h. Nuclei were stained with DAPI and coverslips were mounted with Vectashield medium (Vector Laboratories, Cambridgeshire, UK). Results are presented as a representative example of three biologically independent repetitions of the experiment.

Taminiau A., Draime A., Tys J., Lambert B., Vandeputte J., Nguyen N., Renard P., Geerts D, & Rezsöhazy R. (2016). HOXA1 binds RBCK1/HOIL-1 and TRAF2 and modulates the TNF/NF-κB pathway in a transcription-independent manner. Nucleic Acids Research, 44(15), 7331-7349.

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In Vitro Phosphorylation of GST-GluA2

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Example 4

The purified GST-GluA2 was submitted to in vitro phosphorylation in the presence of the active Src protein (14-326, Merck™) and ATP, for 1 h at 25° C. All reactions were carried out in kinase assay buffer (20 mM MES pH 6.5, 150 mM NaCl, 10 mM MgCl₂, 1mMDTT). A reaction containing GST-GluA2 but without Src kinase was carried out as a negative control. Tyrosine phosphorylation of GST-GluA2 was checked by western blotting using anti-pTyr antibody (PY99) HRP (sc-7020 HRP, Santa Cruz Biotechnology™). Then, the membranes were reprobed with anti-GST antibodies (G7781, Sigma-Aldrich™)

US11834518B2. Interference peptides as inhibitors of interactions related to AMPAR endocytosis (2023-12-05). None [RO], None [RO], None [RO]. Inventors: Stefan Eugen Szedlacsek [RO], Rodica-Aura Badea [RO], Horea-Stefan Szedlacsek [RO], Lucian Hritcu [RO].

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Antibody sources and applications

Cited 3 times

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The antibodies listed in the Key Resources Table were used as follows: rabbit polyclonal antibody to actin (A2066, Sigma-Aldrich, WB 1:2,000), mouse monoclonal to cytochrome c (6H2.B4, 612302, BioLegend, IF 1:200), rabbit polyclonal to DsRed (632496, Clontech Laboratories, IF 1:500, WB 1:1,000), mouse monoclonal to GFP (9F9.F9, ab1218, abcam, IF 1:400–1,000), rabbit polyclonal to GFP (A11122, Thermo Fischer Scientific, IF 1:400), rabbit polyclonal to GFP (ab6556, abcam, WB 1:1,000), rabbit polyclonal to GST (G7781, Sigma-Aldrich, WB 1:20,000), mouse monoclonal to HA (HA-7, H9658, Sigma-Aldrich, IP), rat monoclonal to HA (3F10, 11867423001, Roche, IF 1:100, IP), rabbit polyclonal to optineurin (Ab2, HPA003360, Sigma-Aldrich, IF 1:100), mouse monoclonal to parkin (PRK8, sc-32282, Santa Cruz Biotechnology, WB 1:500), rabbit polyclonal to Tom20 (FL-145, sc-11415, Santa Cruz Biotechnology, IF 1:100), mouse monoclonal to α-tubulin (DM1A, T9025, WB 1:2,000), mouse monoclonal to ubiquitin (P4D1, sc-8017, Santa Cruz Biotechnology, WB 1:1,000), mouse monoclonal to ubiquitin (FK2, ST1200, Millipore, WB 1:1,000). Affinity-purified rabbit polyclonal antibodies against MYO6, TAX1BP1, NDP52, and OPTN (IF 1:100, WB 1:1,000) were generated as previously described (Buss et al., 1998 (link), Morriswood et al., 2007 (link), Sahlender et al., 2005 (link)).

Kruppa A.J., Kishi-Itakura C., Masters T.A., Rorbach J.E., Grice G.L., Kendrick-Jones J., Nathan J.A., Minczuk M, & Buss F. (2018). Myosin VI-Dependent Actin Cages Encapsulate Parkin-Positive Damaged Mitochondria. Developmental Cell, 44(4), 484-499.e6.

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Quantifying γTuSC-Spc110p Interaction

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Our purification method selects for γTuSC that is competent to interact with Spc110p. To quantify the fraction of γTuSC within baculovirus-infected insect cells that is capable of interacting with Spc110p, we performed a series of γTuSC purifications with varying amounts of glutathione-Sepharose resin. The high-speed supernatant and flowthrough fractions were analyzed by Western blotting for each condition. γTuSC^CFP/YFP and GST-Spc110p^1‑220 were detected by α-GFP (1:4000; A-11122; Life Technologies) and α-GST (1:8000; G7781, Sigma-Aldrich), respectively, imaged by fluorophore-conjugated goat α-rabbit secondary antibody (1:2000; 926-68021; Licor Biosciences, Lincoln, NE). Blots were scanned using a Licor Odyssey scanner. Band intensities were quantified using Fiji (Schindelin et al., 2012 (link)).

Lyon A.S., Morin G., Moritz M., Yabut K.C., Vojnar T., Zelter A., Muller E., Davis T.N, & Agard D.A. (2016). Higher-order oligomerization of Spc110p drives γ-tubulin ring complex assembly. Molecular Biology of the Cell, 27(14), 2245-2258.

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GST Pull-Down Assay for Protein-Protein Interactions

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In vitro translated (IVT) proteins were generated with pCS2-N-terminal 6xMYC vectors previously described using TnT® SP6 High-Yield Wheat Germ Extract (L3260, Promega) and by following manufacturer’s recommendations. For each pull-down reaction, 2.5 μg of GST or GST-tagged protein was added along with 1–10 μL IVT protein and 5 μL of washed Glutathione Sepharose 4B beads in 300 uL binding buffer (25 mM HEPES-NaOH [pH 7.4], 500 mM NaCl, 5 mM MgCl₂, and 0.1% Triton X-100) and mixed for 1 hr at 4°C. The beads were washed five times with the same buffer and eluted with LDS sample buffer. The eluates were then resolved by SDS-PAGE and analyzed by immunoblotting with anti-GST (G7781, Sigma-Aldrich) and anti-C-MYC (CBL430, EMD Millipore) antibodies. For pull-down assays using nucleotide-loaded IVT RABL2B, IVT RABL2B proteins were first generated as described above, then incubated at room temperature in the presence of 5 mM MgCl₂ and 1 mM GTP or GDP. The pull-down assays were performed as described with the loaded IVT proteins, with the exception that the binding buffer contained 1 mM GTP or GDP.

Kanie T., Abbott K.L., Mooney N.A., Plowey E.D., Demeter J, & Jackson P.K. (2017). The CEP19-RABL2 GTPase complex binds IFT-B to initiate intraflagellar transport at the ciliary base. Developmental cell, 42(1), 22-36.e12.

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GST Pull-Down Assay with Detailed Antibodies

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GST pull-down was performed with Glutathione Sepharose beads (GE healthcare, Chalffint, UK) and carried out in low-salt EBC lysis buffer (150 mM NaCl; 50 mM Tris, pH 7.5; 1 mM EDTA; 0.5% NP40). Antibodies included in this study: CEP131 (A301-425A, Bethyl, WB 1:1000; ab84864, Abcam, Cambridge, UK, IF 1:300), PCM1 (sc-50164, Santa Cruz, IF 1:200), SSX2IP (HPA027306, Sigma-Aldrich, WB 1:1000), MK2 (3042S, Cell Signalling, Danvers, MA, USA, WB 1:1000), HSP27 pS82 (9709, Cell Signalling, WB 1:5000), GST (G7781, Sigma-Aldrich, WB 1:1000), Pericentrin (ab4448, Abcam, IF 1:1000), p38 MAPK (9212S, Cell Signalling, WB 1:1000), p38 MAPK phospho-Thr180/Tyr182 (9216S, Cell Signalling, WB 1:1000), γ-tubulin (T5326, Sigma-Aldrich, IF 1:250), p150 (610473, BD Biosciences, WB 1:10,000), NOGO-B (previously described by Rousseau et al. [66 (link)] WB 1:1000), SRF phospho-Ser103 (4261, Cell Signalling, WB 1:1000), and α-tubulin (T9026, Sigma-Aldrich, IF 1:500).

Nielsen J.C., Nordgaard C., Tollenaere M.A, & Bekker-Jensen S. (2018). Osmotic Stress Blocks Mobility and Dynamic Regulation of Centriolar Satellites. Cells, 7(7), 65.

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Antibody Immunoblotting Protocol

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GFP antibody (1:5,000; G1544), GST antibody (1:5,000; G7781), and beta-actin (1:10,000; A5441) were purchased from Sigma Aldrich (Sant Luis, USA). Secondary antibodies coupled to horseradish peroxidase used for immunoblotting were obtained from Sigma (A4914 Goat anti-Rabbit IgG).

Roig S.R., Thijssen N., Erp M.v., Ginsel L., Hoenderop J.G.J., & Wijst J.v.d. (2021). Functional basis for calmodulation of the TRPV5 calcium channel.

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