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Gst hrp

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The GST-HRP is a laboratory reagent that consists of Glutathione S-Transferase (GST) fused to Horseradish Peroxidase (HRP). It is a tool used in various biochemical and immunological applications that require the detection and quantification of target proteins or molecules.

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Lab products found in correlation

Flag hrp, by Merck Group (3 mentions) Ccnd2, by Cell Signaling Technology (2 mentions) Cdkn1b, by Cell Signaling Technology (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) γ tubulin, by Merck Group (2 mentions) α tubulin, by Merck Group (2 mentions) Ha and myc, by Santa Cruz Biotechnology (2 mentions) Tomm20, by Cell Signaling Technology (1 mentions) Erp57, by Cell Signaling Technology (1 mentions) Aβ1 42 elisa kit, by Thermo Fisher Scientific (1 mentions) At100, by Thermo Fisher Scientific (1 mentions) Histone h3, by Cell Signaling Technology (1 mentions) Lamp1, by Santa Cruz Biotechnology (1 mentions) β actin, by Cell Signaling Technology (1 mentions) Ha and gfp, by Santa Cruz Biotechnology (1 mentions) β actin, by Merck Group (1 mentions) Fbxw7, by Abcam (1 mentions) Ha hrp, by Merck Group (1 mentions) Goat anti mouse igg hrp, by Jackson ImmunoResearch (1 mentions) Goat anti rabbit igg hrp, by Jackson ImmunoResearch (1 mentions)

Spelling variants of this product

HRP-conjugated anti-GST antibodies (2 citations) Anti-GST-HRP (DG122-2A7, (2 citations)

9 protocols using gst hrp

1

Western Blotting Antibody Deployment

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The following antibodies were used for the western blotting: MYC (Cell signaling 5605S, 1:1000 dilution), γ-tubulin (Sigma T-5326, 1:2000 dilution), Flag-HRP (Sigma A8592, 1:1000 dilution), GST-HRP (Sigma A7340, 1:1000 dilution), MKK3 (Santa Cruz sc-961, 1:1000 dilution), MYC (Cell signaling 5605S, 1:1000 dilution), GAPDH (Cell Signaling 2118, 1:1000 dilution), CDKN1B (p27, Cell Signaling 2552, 1:1000 dilution), CCND2 (Cell Signaling 3741P, 1:1000 dilution), CDK4 (Santa Cruz sc-260, 1:1000 dilution).
Ivanov A.A., Gonzalez-Pecchi V., Khuri L.F., Niu Q., Wang Y., Xu Y., Bai Y., Mo X.L., Prochownik E.V., Johns M.A., Du Y., Khuri F.R, & Fu H. (2017). OncoPPi-informed discovery of Mitogen-Activated Protein Kinase Kinase 3 as a novel binding partner of c-Myc. Oncogene, 36(42), 5852-5860.
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2

Characterization of APP and Tau Fragments

Cited 2 times
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Primary antibodies to the following targets were used: APP N585, APP C586 and Tau N368 which specifically recognize the δ-secretase-derived APP and tau fragments respectively, were described previously(Zhang et al., 2014 (link); Zhang et al., 2015 (link)). AEP 6E3 and 11b7 (from Dr Colin Watts, University of Dundee), GST-HRP (Sigma-Aldrich, # GE RPN1236), LAMP1 (Santa Cruz, #5570), GFP (Santa Cruz, #101525), TrkB (Santa Cruz, #377218), myc (Santa Cruz, #M4439), sAPPβ (6A1, IBL, #10321), AT100 (Thermo, #MN1060), BACE1 N terminal (abcam, #ab79921), APP N terminal (22C11, Calbiochem, #MAB348), Aβ 4G8 (Signet, #800709), β-actin (Cell Signaling, #3700), BACE1 C terminal (Cell Signaling, #5606), EEA1 (Cell Signaling, #3288), GGA3 (Cell Signaling, #8027), Histone H3 (Cell Signaling, #4499), TOMM 20 (Cell Signaling, #42406), ERp57 (Cell Signaling, #2881), see details in Table S2. Mouse and human Aβ1–40 and Aβ1–42 ELISA kits were purchased from Invitrogen, recombinant AEP was purchased from Novoprotein.
Xia Y., Wang Z.H., Zhang Z., Liu X., Yu S.P., Wang J.Z., Wang X.C, & Ye K. (2021). Delta- and Beta- Secretases Crosstalk Amplifies the Amyloidogenic Pathway in Alzheimer’s disease. Progress in neurobiology, 204, 102113.
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3

Protein Extraction and Western Blot Analysis

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The mouse brain tissue or human tissue samples were lysed in lysis buffer (50 mM Tris, pH 7.4, 40 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1.5 mM Na3VO4, 50 mM NaF, 10 mM sodium pyrophosphate, 10 mM sodium β-glycerophosphate, supplemented with a cocktail of protease inhibitors), and centrifuged for 15 min at 16,000 × g. The supernatant was boiled in SDS loading buffer. After SDS-PAGE, the samples were transferred to a nitrocellulose membrane. Primary antibodies to the following targets were used: GST-HRP, α-tubulin, and β-actin (Sigma-Aldrich), HA and GFP (Santa Cruz Biotechnology), BACE1, STAT1, p-STAT1, JAK2, p-JAK2, and p-SGK1 (cell signaling), SGK1 (Abcam), HT7 and AT8 (Thermo Fisher), δ-secretase antibody (clone 6E3, president from Dr. Colin Watts, University of Cambridge).
Zhang Z., Li X.G., Wang Z.H., Song M., Ping Yu S., Su Kang S., Liu X., Zhang Z., Xie M., Liu G.P., Wang J.Z, & Ye K. (2018). δ-Secretase-cleaved Tau stimulates Aβ production via upregulating STAT1-BACE1 signaling in Alzheimer’s disease. Molecular psychiatry, 26(2), 586-603.
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4

Protein extraction and immunoblotting

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The mouse brain tissue or human tissue samples were lysed in lysis buffer (50 mM Tris, pH 7.4, 40 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1.5 mM Na3VO4, 50 mM NaF, 10 mM sodium pyrophosphate and 10 mM sodium β-glycerophosphate, supplemented with a cocktail of protease inhibitors), and centrifuged for 15 min at 16,000g. The supernatant was boiled in SDS loading buffer. After SDS–PAGE, the samples were transferred to a nitrocellulose membrane. Primary antibodies to the following targets were used: GST-HRP (Sigma-Aldrich, GERPN1236), α-tubulin (Sigma-Aldrich, T9026), HA (Santa Cruz, H3663), myc (Santa Cruz, M4439), α-synuclein N terminus (Santa Cruz, sc-514908); anti-α-synuclein (syn1, clone 42, BD Bioscience, 610787), AEP (clone 6E3, gift from C. Watts), TH (Cell Signaling Technology, 2792), α-synuclein C terminus (Cell Signaling Technology, 2642) and α-synuclein LB509 (Thermo Fisher Scientific, 180215).
Zhang Z., Kang S.S., Liu X., Ahn E.H., Zhang Z., He L., Iuvone P.M., Duong D.M., Seyfried N.T., Benskey M.J., Manfredsson F.P., Jin L., Sun Y.E., Wang J.Z, & Ye K. (2017). Asparagine endopeptidase cleaves α-synuclein and mediates pathologic activities in Parkinson’s disease. Nature structural & molecular biology, 24(8), 632-642.
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5

Western Blotting Antibody Deployment

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The following antibodies were used for the western blotting: MYC (Cell signaling 5605S, 1:1000 dilution), γ-tubulin (Sigma T-5326, 1:2000 dilution), Flag-HRP (Sigma A8592, 1:1000 dilution), GST-HRP (Sigma A7340, 1:1000 dilution), MKK3 (Santa Cruz sc-961, 1:1000 dilution), MYC (Cell signaling 5605S, 1:1000 dilution), GAPDH (Cell Signaling 2118, 1:1000 dilution), CDKN1B (p27, Cell Signaling 2552, 1:1000 dilution), CCND2 (Cell Signaling 3741P, 1:1000 dilution), CDK4 (Santa Cruz sc-260, 1:1000 dilution).
Ivanov A.A., Gonzalez-Pecchi V., Khuri L.F., Niu Q., Wang Y., Xu Y., Bai Y., Mo X.L., Prochownik E.V., Johns M.A., Du Y., Khuri F.R, & Fu H. (2017). OncoPPi-informed discovery of Mitogen-Activated Protein Kinase Kinase 3 as a novel binding partner of c-Myc. Oncogene, 36(42), 5852-5860.
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6

Western Blot Analysis of Brain Proteins

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The mouse brain tissue or human tissue samples was lysed in lysis buffer (50 mM Tris, pH 7.4, 40 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1.5 mM Na3VO4, 50 mM NaF, 10 mM sodium pyrophosphate, 10 mM sodium β-glycerophosphate, supplemented with protease inhibitors cocktail), and centrifuged for 15 min at 16,000 g. The supernatant was boiled in SDS loading buffer. After SDS-PAGE, the samples were transferred to a nitrocellulose membrane. Primary antibodies to the following targets were used: GST-HRP (Sigma-Aldrich), tubulin (Sigma-Aldrich), HA and myc (both from Santa Cruz), His (GE healthcare), HT7, tau5, AT8 and AT100 (Thermo), and tau-1 (Calbiochem).
Zhang Z., Song M., Liu X., Kang S.S., Kwon I.S., Duong D.M., Seyfried N.T., Hu W.T., Liu Z., Wang J.Z., Cheng L., Sun Y.E., Yu S.P., Levey A.I, & Ye K. (2014). Cleavage of tau by asparagine endopeptidase mediates the neurofibrillary pathology in Alzheimer’s disease. Nature medicine, 20(11), 1254-1262.
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7

Rpb4/Rpb7-HA Immunoprecipitation and GST-Ssu72 Binding

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Cells were grown to an OD600 of 0.8, collected, washed and suspended in lysis buffer (20 mM HEPES pH 7.6, 200 mM KCH3COO−, 1 mM EDTA pH 8.0, glycerol 10%) with protease and phosphatase inhibitors. Yeast whole-cell extracts were prepared by glass bead disruption of cells using a FastPrep system. Protein concentrations were determined and 5 mg of total protein was incubated with 1.5 μl anti-HA-Sepharose beads for 2 h at 4°C to immunoprecipitate Rpb4/Rpb7-HA. The immunoprecipitates were washed several times with lysis buffer and then incubated with 2 μg of either GST or GST-Ssu72 proteins for 90 min at 4°C. Reaction mixtures were extensively washed and afterward electrophoresed on a SDS-polyacrylamide gels, transferred and immunoblotted with the following antibodies: anti-HA, anti-Rpb4 and GST-HRP (A3740, Sigma).
Allepuz-Fuster P., O’Brien M.J., González-Polo N., Pereira B., Dhoondia Z., Ansari A, & Calvo O. (2019). RNA polymerase II plays an active role in the formation of gene loops through the Rpb4 subunit. Nucleic Acids Research, 47(17), 8975-8987.
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8

Western Blot Analysis of Brain Proteins

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The mouse brain tissue or human tissue samples was lysed in lysis buffer (50 mM Tris, pH 7.4, 40 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1.5 mM Na3VO4, 50 mM NaF, 10 mM sodium pyrophosphate, 10 mM sodium β-glycerophosphate, supplemented with protease inhibitors cocktail), and centrifuged for 15 min at 16,000 g. The supernatant was boiled in SDS loading buffer. After SDS-PAGE, the samples were transferred to a nitrocellulose membrane. Primary antibodies to the following targets were used: GST-HRP (Sigma-Aldrich), tubulin (Sigma-Aldrich), HA and myc (both from Santa Cruz), His (GE healthcare), HT7, tau5, AT8 and AT100 (Thermo), and tau-1 (Calbiochem).
Zhang Z., Song M., Liu X., Kang S.S., Kwon I.S., Duong D.M., Seyfried N.T., Hu W.T., Liu Z., Wang J.Z., Cheng L., Sun Y.E., Yu S.P., Levey A.I, & Ye K. (2014). Cleavage of tau by asparagine endopeptidase mediates the neurofibrillary pathology in Alzheimer’s disease. Nature medicine, 20(11), 1254-1262.
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9

Western Blotting Antibody Specifications

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The following antibodies and conjugates were used for the western blotting: Flag-HRP (Sigma, Cat# A8592, dilution 1:2000), GST-HRP (Sigma, Cat# A7340, dilution 1:3000), HA-HRP (Sigma, Cat# H6533, dilution 1:3000), goat anti-mouse IgG-HRP (Jackson ImmunoResearch, Cat# 115-035-003, dilution 1:3000), and goat anti-rabbit IgG-HRP (Jackson ImmunoResearch, Cat# 111-035-003, dilution 1:3000), and antibodies for MYC (Cell Signaling Technology, Cat# 5605S, dilution 1:1000), MYC (Santa Cruz, Cat# sc-40, dilution 1:1000), FBXW7 (Abcam, Cat# ab109617, dilution 1:1000), ERK (Cell Signaling Technology, Cat# 4695, dilution 1:1000), GSK3β (Cell Signaling Technology, Cat# 9315, dilution 1:1000), actin (Sigma, Cat# A5441, dilution 1:3000), GFP (Abcam, Cat# ab290, 1:6000), and Hsp60 (Cell Signaling Technology, Cat# 4870, dilution 1:1000).
Gonzalez-Pecchi V., Kwan A.K., Doyle S., Ivanov A.A., Du Y, & Fu H. (2019). NSD3S stabilizes MYC through hindering its interaction with FBXW7. Journal of Molecular Cell Biology, 12(6), 438-447.
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