Goat anti mouse hrp

Manufactured by Promega

Sourced in United Kingdom, United States

Goat-anti-mouse-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and bind to mouse primary antibodies in immunoassays and other immunochemical applications.

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Lab products found in correlation

Goat anti rabbit hrp, by Promega (4 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (3 mentions) Lsm 880 confocal microscope, by Zeiss (2 mentions) Immobilon p 0.45 m pvdf membrane, by Merck Group (1 mentions) Western lightening plus ecl kit, by PerkinElmer (1 mentions) Gradient sds page gel, by Bio-Rad (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Goat anti rat hrp, by Jackson ImmunoResearch (1 mentions) Anti gfp sc 8334, by Santa Cruz Biotechnology (1 mentions) Protein assay dye reagent concentrate, by Bio-Rad (1 mentions) Anti flag m2, by Merck Group (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Anti ha 3f10, by Roche (1 mentions) Trans blot apparatus, by Bio-Rad (1 mentions) Mgme1, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Ab10436, by Abcam (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Sds page 4 12 bis tris gels, by Thermo Fisher Scientific (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Goat anti-mouse IgG (H+L) HRP HRP-conjugated goat anti-mouse IgG (H+L) HRP-conjugated goat anti-mouse Goat anti-mouse IgG-HRP HRP-conjugated goat anti-mouse secondary antibody HRP-conjugated goat anti-mouse or anti-rabbit secondary antibody Goat anti-mouse HRP-conjugated secondary antibodies Goat anti-rabbit and goat anti-mouse HRP-conjugated secondary antibodies Goat anti-mouse IgG HRP conjugate HRP)-conjugated goat anti-mouse antibody

6 protocols using goat anti mouse hrp

Western Blot Protocol for CBX7 and DCAF12l1 Quantification

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20µl of protein extracts were resolved on 4%–20% gradient SDS-PAGE gels (Bio-Rad) and proteins were transferred for 1hr on 100V in transfer buffer (48mM Tris, 39mM Glycine, 20% methanol) to Immobilon-P 0.45µm PVDF membrane (Millipore) using Mini Protean Tetra transfer unit (Bio-Rad)). To detect CBX7 protein expression, Western blotting was performed with mouse monoclonal CBX7 Antibody (G-3) (Santa Cruz Biotechnologies, sc-376274) as primary antibody and goat-anti-mouse-HRP (Promega) as a secondary antibody. For quantitative Western blotting of DCAF12l1 protein, anti-WDR40B (Dcaf12l1) rabbit polyclonal antibody (Biorbit, orb155395) was used as a primary antibody along with anti-Ctcf rabbit polyclonal antibody (Cell Signaling Technologies, #2899) as a loading control. Goat-anti-rabbit- HRP (Promega) was employed as a secondary antibody. Protein bands were developed using Western Lightening Plus -ECL Kit (Perkin-Elmer) and the signal intensity was analyzed using Chemidoc MP Imaging System (Bio-Rad) and ImageLab Ver. 5.2.1 software (Bio-Rad). Exposures were captured on different times using ChemiDoc cumulative signal option to avoid signal saturation. Standard curves were prepared using increasing amounts of cell extract (Fig. 6 G), to confirm a signal intensity staying in a dynamic linear range.

Rosenberg M., Blum R., Kesner B., Maier V.K., Szanto A, & Lee J.T. (2017). Denaturing CLIP (dCLIP) pipeline identifies discrete RNA footprints on chromatin-associated proteins and reveals that CBX7 targets 3’UTRs to regulate mRNA expression. Cell systems, 5(4), 368-385.e15.

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Monitoring p53 Stability in Transfected Cells

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H1299 cells were transfected with 1.5 μg of p53 expression plasmids and 250 ng pEGFP-N1 (transfection control) using Lipofectamine3000 according to the manufacturer’s protocol and harvested 18 hours after transfection. To monitor potential differences of p53 stability, transfected cells were treated with 5 μM MG132 8–12 hours prior to harvesting. After treatment or transfection cells were collected, washed in PBS and lysed for 30 min on ice in RIPA buffer (20 mM Tris pH 7.5, 150 mM NaCl, 1 mM Na₂EDTA, 1 mM EGTA, 1% NP-40, 1% sodium deoxycholate) including 1x cOmplete Protease Inhibitor Cocktail (Roche). Extracts obtained from the transactivation assay were used without adding additional inhibitors. Lysates were centrifuged for 30 min at 4°C and 17,000 g and supernatants were quantified using Bio-Rad Protein Assay Dye Reagent Concentrate (Bio-Rad). Thirty μg of denatured cell lysate/lane were loaded to 12% SDS-PAGE. After blotting, proteins were detected with anti-HA (3F10, Roche), anti-GFP (sc-8334, Santa Cruz), anti-FLAG M2 (F3165, Sigma-Aldrich) or anti-mouse-actin antibodies prior to detection with goat anti-mouse-HRP (W402B, Promega) or goat anti-rat-HRP (Jackson ImmunoResearch, Newmarket, UK).

Hasche D., Stephan S., Braspenning-Wesch I., Mikulec J., Niebler M., Gröne H.J., Flechtenmacher C., Akgül B., Rösl F, & Vinzón S.E. (2017). The interplay of UV and cutaneous papillomavirus infection in skin cancer development. PLoS Pathogens, 13(11), e1006723.

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Western Blotting of Immune Receptors

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Western blotting was performed as described in Bonnin et al. [2 (link)], with the following modifications. Proteins were transferred onto PVDF membranes using semi-dry Trans-Blot apparatus (Bio-Rad), anti-actin was from MP (clone C4, France; RRID:AB_2335127), anti-hTLR3 (clone D10F10; RRID:AB_10829166; MA, USA), anti-hTLR7 (clone D7; RRID:AB_10692895), anti-hTLR8 (clone D3Z6J; RRID:AB_2797755), anti-hTLR9 (clone D9M9H; RRID:AB_2798290), anti-RIG-I (clone D14G6; RRID:AB_2269233), anti-MDA5 (clone D74E4; RRID:AB_10694490) were from Cell Signaling, and secondary Goat anti-Rabbit HRP (RRID:AB_430833) conjugate and secondary Goat anti-Mouse HRP (RRID:AB_430834) conjugate were from Promega. Image acquisition and relative band intensity measurement were performed using a Chemi Doc XR+ apparatus (Bio-Rad) driven by Image Lab software version 6.1.

Thierry S., Maadadi S., Berton A., Dimier L., Perret C., Vey N., Ourfali S., Saccas M., Caron S., Boucard-Jourdin M., Colombel M., Werle B, & Bonnin M. (2023). TL-532, a novel specific Toll-like receptor 3 agonist rationally designed for targeting cancers: discovery process and biological characterization. Microbial Cell, 10(6), 117-132.

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Western Blot Analysis of Mitochondrial Proteins

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Primary antibodies used for western blotting were as follows: MGME1 (Sigma HPA040913, 1 : 500 dilution), β-actin (Sigma A2228, 1 : 300 000), PolgA (Santa Cruz, 1 : 1000), mtSSB1 (kindly donated by Dr Kang) and TFAM (kindly donated by Dr Wiesner). Secondary antibodies used were as follows: goat anti-rabbit HRP (Promega W401B, 1 : 2000) and goat anti-mouse HRP (Promega W402B, 1 : 2000).

Nicholls T.J., Zsurka G., Peeva V., Schöler S., Szczesny R.J., Cysewski D., Reyes A., Kornblum C., Sciacco M., Moggio M., Dziembowski A., Kunz W.S, & Minczuk M. (2014). Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease. Human Molecular Genetics, 23(23), 6147-6162.

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Immunofluorescence and Western Blot Analyses

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Localisation of proteins by immunofluorescence was carried out in fixed 143B cells as previously described (22 (link)). Images were captured using a Zeiss LSM 880 confocal microscope.
The following antibodies were used for immunofluorescence experiments in this work: mouse anti-TOM22 (Abcam, ab10436, 1:250), Alexafluor-594 anti-mouse (Molecular Probes, A11005, 1:1000), rabbit anti-TFAM (gifted by Prof. Rudolf Wiesner, 1:500), Alexafluor-405 anti-rabbit (Molecular Probes, A31553, 1:1000), rat anti-HA (Roche, 11867431001, 1:500), Alexafluor-488 anti-rat (Molecular Probes, A11006, 1:1000). Mounting medium used was either ProLong Gold Antifade Mountant (Molecular Probes), or ProLong Gold Antifade Mountant with DAPI (Molecular Probes).
For western blot analyses, ∼20 μg of extracted proteins were resolved on SDS-PAGE 4–12% bis-tris gels (Life Technologies). The following antibodies were used for western blotting in this work: mouse anti-FLAG (Sigma, F1804, 1:2000), rabbit anti-FLAG (Sigma, F7425, 1:2000), rat anti-HA (Roche, 11867431001, 1:1000), rabbit anti-Histone H4 (Abcam, ab10158, 1:5000), rabbit anti-SSB1 (kindly gifted by Prof D. Kang, 1:4000), mouse anti-TOM22 (Abcam, ab10436, 1:5000), mouse anti-GAPDH (Abcam, ab9484, 1:10 000), goat anti-rabbit HRP (Promega, W401B, 1:2000), goat anti-mouse HRP (Promega, W402B, 1:2000), goat anti-rat HRP (Santa Cruz, SC2065, 1:1000).

Gammage P.A., Gaude E., Van Haute L., Rebelo-Guiomar P., Jackson C.B., Rorbach J., Pekalski M.L., Robinson A.J., Charpentier M., Concordet J.P., Frezza C, & Minczuk M. (2016). Near-complete elimination of mutant mtDNA by iterative or dynamic dose-controlled treatment with mtZFNs. Nucleic Acids Research, 44(16), 7804-7816.

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Western Blot Analysis of Liver Samples

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Liver samples were lysed in RIPA buffer (Cell Signaling Technology) supplemented with protease and phosphatase inhibitors. Protein concentration was determined by using the PierceTM BCA Protein Assay Kit (Thermo Scientific). Lysates were separated by SDS-PAGE and transferred to a Whatman Protran BA85 membrane (GE Healthcare). Membranes were incubated with the following primary antibodies overnight: Brg1 (Santa Cruz Biotechnology), β-tubulin (Abcam), β-actin(Santa Cruz Biotechnology), PCNA (Cell Signaling Technology, Inc.), pH3 (Cell Signaling Technology, Inc.), Cyclin B1 (Cell Signaling Technology, Inc.), Cdk1 (Cell Signaling Technology, Inc.), GAPDH (Santa Cruz Biotechnology), p53 (Leica Biosystems), then incubated with Secondary antibody goat-anti-rabbit-HRP or goat-anti-mouse-HRP (Promega) for 1 h. Antibody binding was visualized using the Pierce™ ECL western blotting detection system (GE Healthcare). Densitometric analysis was performed using the ImageJ software (https://imagej.nih.gov/ij/).

Wang B., Kaufmann B., Engleitner T., Lu M., Mogler C., Olsavszky V., Öllinger R., Zhong S., Geraud C., Cheng Z., Rad R.R., Schmid R.M., Friess H., Hüser N., Hartmann D, & von Figura G. (2019). Brg1 promotes liver regeneration after partial hepatectomy via regulation of cell cycle. Scientific Reports, 9, 2320.

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