Anti rgs his

Manufactured by Qiagen

The Anti-RGS-His is a lab equipment product that detects the presence of RGS-His proteins. It is a crucial tool for researchers studying cellular signaling pathways involving RGS-His proteins.

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Lab products found in correlation

Anti gfp, by Proteintech (3 mentions) Anti ha, by Roche (3 mentions) Anti myc, by Proteintech (3 mentions) Anti vinculin, by Merck Group (2 mentions) Anti ubiquitin, by Agilent Technologies (2 mentions) Anti gapdh, by Cell Signaling Technology (2 mentions) Anti β actin, by Merck Group (2 mentions) Anti flag, by Merck Group (2 mentions) Anti α tubulin, by Merck Group (2 mentions) Anti na k atpase α 1, by Merck Group (1 mentions) Anti rat hrp, by Thermo Fisher Scientific (1 mentions) Anti pma1, by Abcam (1 mentions) Anti hsp70, by Thermo Fisher Scientific (1 mentions) Anti mouse hrp, by Agilent Technologies (1 mentions) Anti rabbit hrp, by Agilent Technologies (1 mentions) Anti α tubulin, by Abcam (1 mentions) Anti hdac6, by Cell Signaling Technology (1 mentions) Anti tubulin, by Merck Group (1 mentions) 0.2 μm nitrocellulose membranes, by Advantec (1 mentions) Amersham ecl detection reagent, by GE Healthcare (1 mentions)

6 protocols using anti rgs his

Western Blot Analysis of Cellular Proteins

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SDS-PAGE was performed using 8% and 12.5% acrylamide gels. For the Western blotting procedure, 0.2 μm nitrocellulose membranes were used. Following protein transfer to membranes, these were blocked in 5% fat-free milk powder, 5 mM NaN₃ and 0.1% Tween-20 in PBS.
Antibodies and their sources were: anti-HA (Roche, 15645900), anti-Cdc48 (own production), anti-α-tubulin (Abcam, YL1/2 MA1-80017), anti-Pma1 (Abcam, 40B7 Ab4645), anti-RGS-His (Qiagen, 34610), anti-Vinculin (Sigma, hVIN1 V9264), anti-GAPDH (Cell signalling technology, 14C10 2118), anti-Na/K ATPase α-1 (Merck, C464.6 05–369), anti-Hsp105 (HSPH1, Abcam, Ab108625), anti-HSPA4 (Abcam, Ab185219), anti-Ubiquitin (Dako, Z0458), anti-Myc (Chromotek, 9E1 9e1-100), anti-Rpn1 (Enzo Life Sciences, p112-1 PW9270), anti-20S α-subunits (Enzo Life Sciences, MCP231 PW8195), anti-Hsp70 (Invitrogen, 5A5 MA3-007), anti-Aspartoacylase (Thermo Scientific, PA5-29180), anti-GFP (Chromotek, 3H9 3h9-100). Secondary antibodies and their sources were: HRP-anti-rat (Invitrogen, 31470), HRP-anti-mouse (Dako, P0260), HRP-anti-rabbit (Dako, P0448).

Gersing S.K., Wang Y., Grønbæk-Thygesen M., Kampmeyer C., Clausen L., Willemoës M., Andréasson C., Stein A., Lindorff-Larsen K, & Hartmann-Petersen R. (2021). Mapping the degradation pathway of a disease-linked aspartoacylase variant. PLoS Genetics, 17(4), e1009539.

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Western Blot Analysis of DNA Repair Proteins

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Proteins were separated on 7 cm x 8 cm 8% acrylamide gels and subsequently transferred to 0.2 μm nitrocellulose membranes. Membranes were blocked in PBS containing 5% fat-free milk powder and 0.1% Tween-20. Membranes were then probed with the indicated antibodies overnight.
Antibodies and their sources were: anti-human MSH2 (CalBiochem), anti-human MSH6 (BD Biosciences), anti-human MSH3 (Abcam), anti-RGSHis (Qiagen), anti-HDAC6 (Cell Signaling Technology), anti-tubulin (Sigma) and anti-β-actin (Sigma). All secondary antibodies were purchased from DAKO.

Nielsen S.V., Stein A., Dinitzen A.B., Papaleo E., Tatham M.H., Poulsen E.G., Kassem M.M., Rasmussen L.J., Lindorff-Larsen K, & Hartmann-Petersen R. (2017). Predicting the impact of Lynch syndrome-causing missense mutations from structural calculations. PLoS Genetics, 13(4), e1006739.

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Western Blot Analysis with SDS-PAGE Separation

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SDS-PAGE was performed with either 8% or 12.5% acrylamide gels. For the subsequent Western blotting, 0.2 μm nitrocellulose membranes (Advantec, Toyo Roshi Kaisha Ltd.) were used. The membranes were blocked in PBS containing 5% fat-free milk powder, 0.1% Tween-20 and 5 mM NaN₃ for at least 30 min. Incubation with the primary antibodies, all diluted in PBS containing 5% BSA and 0.1% Tween-20, was performed overnight at 4 °C. Primary antibodies used for this study were: anti-HA (Roche, 1:3000, 11867423001), anti-RGS-His (Qiagen, 1:2000, 34610), anti-Myc (ChromoTek, 1:1000, AB_2631398), anti-Vinculin (Sigma, 1:2000, V9264), anti-α-tubulin (Merck, 1:1000, TAT-1 00020911), anti-GFP (ChromoTek, 1:1000, 3H9), anti-RFP (ChromoTek, 1:1000, 6G6). The secondary antibodies were: HRP-antimouse IgG (Dako, 1:5000, P0260), HRP-antirat IgG (Thermo Fisher Scientific, 1:5000, 31470). The blots were developed using Amersham ECL detection reagent (GE Healthcare) and a ChemiDoc Imaging System (BioRad).

Kampmeyer C., Grønbæk-Thygesen M., Oelerich N., Tatham M.H., Cagiada M., Lindorff-Larsen K., Boomsma W., Hofmann K, & Hartmann-Petersen R. (2023). Lysine deserts prevent adventitious ubiquitylation of ubiquitin-proteasome components. Cellular and Molecular Life Sciences, 80(6), 143.

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Western Blotting Optimization Protocol

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Unless otherwise stated whole cell extracts were prepared by lysing cells directly in SDS sample buffer. SDS-PAGE was performed on 12.5% acrylamide gels. For Western blotting 0.2 μm nitrocellulose membranes were used. For blocking, the membranes were incubated in 5% fat-free milk powder and 0.1% Tween-20 in PBS. Antibodies and their sources were: anti-ubiquitin (Z0458, DAKO), anti-Rpt6 (p45-110, Enzo Biosciences), anti-20S proteasome α-subunits (MCP231, Enzo Biosciences), anti-FLCN (D14G9, Cell Signaling Technology), anti-GFP (3H9, Chromotek), anti-myc (9E1, Chromotek) anti-HA (12CA5, Roche), anti-Flag (M2, Sigma), anti-RGSHis (34650, Qiagen), anti-β-actin (AC74, Sigma), anti-NaK-ATPase (C464.6, Sigma), anti-LC3A/B (D3U4C, Cell Signaling Technology), anti-GAPDH (14C10, Cell Signaling Technology), anti-α tubulin (TAT-1, 00020911 Sigma). All secondary antibodies were purchased from DAKO. The Un-Scan-It v6.1 software (Silk Scientific) was used for densitometry.

Clausen L., Stein A., Grønbæk-Thygesen M., Nygaard L., Søltoft C.L., Nielsen S.V., Lisby M., Ravid T., Lindorff-Larsen K, & Hartmann-Petersen R. (2020). Folliculin variants linked to Birt-Hogg-Dubé syndrome are targeted for proteasomal degradation. PLoS Genetics, 16(11), e1009187.

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Generation and characterization of Bmi1 variants

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Flag-Bmi1 (WT or K88R) and Flag-GFP-Bmi1 (WT or K88R) were generated using standard cloning procedures. K88R mutation was introduced into Bmi1 by site directed mutagenesis using QuikChange Site-Directed Mutagenesis Kit (Agilent California, USA). HA-SUMO1, HA-UBC9, RGS-SENP1 and RGS-SENP1m were previously described45 (link)48 (link). Anti-FLAG (mouse; #F1804) and anti-HA(mouse; #H3663) antibodies were from Sigma; anti-RGS-his (mouse; #34610) antibody from QIAGEN; anti-H-Ras (mouse; #sc-53959) and p16^Ink4a (mouse; #sc-1661) antibodies from Santa Cruz; anti-SUMO1 (rabbit; #ab5316), H3K27me3 (mouse; #ab6002) and p19^Arf (rabbit; #ab80) antibodies from Abcam; anti-Bmi1 (mouse; #05–637), γH2AX (mouse; #05–636) and uH2AK119 (mouse; #05–678) antibodies from Millipore.

Xia N., Cai J., Wang F., Dong B., Liu S., Chen F., Cheng J, & Zuo Y. (2016). SENP1 Is a Crucial Regulator for Cell Senescence through DeSUMOylation of Bmi1. Scientific Reports, 6, 34099.

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Western Blot Analysis of Protein Expression

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Whole-cell lysates, prepared from 1 × 10 6 cells with urea lysis buffer (8 mol/L urea, 0.5% Triton X-100, 10 mmol/L dithiothreitol) supplemented with protease inhibitors (Mini Complete Cocktail Tablet, Roche Diagnostics) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane (Amersham Protran, 0.45 μm). Membranes were incubated overnight at 4°C with primary antibodies diluted in 5% bovine serum albumin/1 × Tris-buffered saline (TBS). Primary specific antibodies were purchased from Santa Cruz Biotechnology (anti-SZF1/ZNF589 S-14 sc-100263 and E-17 sc-100261), Abcam (anti-SZF1/ZNF589 ab121173 and ab179541), Cell Signaling Technologies (anti-BCL2 cs-2876, anti-β-actin cs-4970, anti-HIF1α cs-3716, anti-cleaved caspase-3 cs-9661), and Qiagen (anti-RGS-His). After incubation with secondary antibodies (anti-rabbit or anti-mouse horseradish peroxidase, Santa Cruz Biotechnology), blots were washed and signals visualized through chemiluminescence using the Western Lightning Plus-ECL kit from Perkin Elmer.

Venturini L., Stadler M., Manukjan G., Scherr M., Schlegelberger B., Steinemann D, & Ganser A. (2016). The stem cell zinc finger 1 (SZF1)/ZNF589 protein has a human-specific evolutionary nucleotide DNA change and acts as a regulator of cell viability in the hematopoietic system. Experimental hematology, 44(4).

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