Goat anti mouse igg

Manufactured by Abcam

Sourced in United States, United Kingdom, China

Goat anti-mouse IgG is a secondary antibody that binds to primary mouse antibodies. It is used in various immunodetection techniques to amplify and visualize the signal from mouse primary antibodies.

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Lab products found in correlation

Goat anti rabbit igg, by Abcam (46 mentions) Ripa buffer, by Thermo Fisher Scientific (10 mentions) Gapdh, by Abcam (8 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions) Pvdf membrane, by Merck Group (7 mentions) Horseradish peroxidase conjugated goat anti rabbit igg, by Abcam (5 mentions) Goat anti rabbit igg, by Thermo Fisher Scientific (4 mentions) Ripa lysis buffer, by Beyotime (4 mentions) Ab6789, by Abcam (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Quantity one software, by Bio-Rad (3 mentions) Goat anti rat igg, by Abcam (3 mentions) Anti gapdh, by Abcam (3 mentions) Optiblot ecl detection kit, by Abcam (3 mentions) Complete protease inhibitor cocktail, by Roche (3 mentions) Chemidoc, by Bio-Rad (3 mentions) Westernbright quantum, by Advansta (3 mentions) β actin, by Merck Group (3 mentions) Anti β actin, by Abcam (3 mentions) Triton x 100, by Merck Group (2 mentions)

Close spelling variants of this product

HRP-labelled goat anti-mouse IgGs (3 citations) Anti-IgG mouse (2 citations) Mouse anti (2 citations)

126 protocols using goat anti mouse igg

Immunohistochemical Analysis of DUSP2 Expression

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Paraffin-embedded mouse and human kidney sections (2.0-μm-thick) were prepared as described previously 25 (link). A routine Hematoxylin-eosin (H&E) staining of the mouse kidney section was performed in accordance with protocol. DUSP2 expression was quantified using Image Pro-Plus v. 6.0 software (Bethesda, MD, USA).
Immunohistochemical (IHC) staining was performed as described previously 26 (link). Briefly, after de-waxing, hydration, endogenous enzyme, and biotin removal and antigen repair, the sections were incubated with primary antibodies at 4 °C overnight. Then, the Cy3 (Goat Anti-Mouse IgG, Abcam; Goat anti-Rabbit IgG, Invitrogen)- or Alexa Fluor 488-conjugated-conjugated secondary antibody (Goat Anti-Mouse IgG, Abcam; Goat anti-Rabbit IgG, Invitrogen) were incubated for 1 h at room temperature before DAPI (Beyotime Biotechnology, Shanghai, China) staining. PBS was used as a negative control instead of a primary antibody. Slides were photographed using a confocal microscope (ZEISS, LSM780) and analyzed with ZEN 2012 software (version 1.1.1.0, Carl Zeiss Microscopy GmbH).

Xiong J., Ran L., Zhu Y., Wang Y., Wang S., Wang Y., Lan Q., Han W., Liu Y., Huang Y., He T., Li Y., Liu L., Zhao J, & Yang K. (2022). DUSP2-mediated inhibition of tubular epithelial cell pyroptosis confers nephroprotection in acute kidney injury. Theranostics, 12(11), 5069-5085.

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Western Blot Analysis of Protein Targets

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The extraction of cellular proteins and their detection via western blotting were performed as previously described [21 (link), 24 (link)]. Western blotting was performed using the following primary antibodies: mouse anti-AR, mouse anti-beclin1, goat anti-LC3β, mouse anti-LAMP1, and rabbit anti-IKKγ antibodies (1:500, Santa Cruz Biotechnology); mouse anti-TLR4, rabbit anti-iNOS, rabbit anti-IKKα, and rabbit anti-4-HNE (1:1000, Abcam); rabbit anti-IKKβ and rabbit anti-phospho-IκBα antibodies (1:1000, Epitomics); rabbit anti-phospho-IKKα/β, rabbit anti-IκBα, rabbit anti-p65, and rabbit anti-phospho-p65 antibodies (1:1000, Cell Signaling Technology, Danvers, MA, USA); and mouse anti-β-actin antibodies (1:8000, Sigma-Aldrich). The blots were then incubated with their respective secondary antibodies: horseradish peroxidase-conjugated goat anti-rabbit IgG, goat anti-mouse IgG (both 1:8000, Abcam), and donkey anti-goat IgG (1:5000, Santa Cruz Biotechnology). β-actin was used as the loading control. The immunoreactive bands were scanned using the Bio-Rad ChemiDoc™ XRS⁺ imager with Image Lab™ Software (Bio-Rad Laboratories, CA, USA). Band intensity was quantified using Quantity One software (Bio-Rad Laboratories).

Cheng P., Xie J., Liu Z, & Wang J. (2021). Aldose reductase deficiency inhibits LPS-induced M1 response in macrophages by activating autophagy. Cell & Bioscience, 11, 61.

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Analyzing MMP13 and Pol II Ser5P in TNF-α-Stimulated Human NP Cells

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Human NP cells were seeded on coverslips, exposed to 50 ng/ml TNF-ɑ, and treated with or without 50 nM THZ1, 4% paraformaldehyde was used to fix cells for 10 min, 0.1% Triton X-100 permeabilization for 10 min, 10% goat serum block for 1 h, and PBS was used to wash cells three times at intervals before incubation with MMP13 (1:3,000 dilution; ab39012; Abcam), Pol II Ser5P (1:1,000 dilution; ab5408; Abcam) at 4°C overnight. After washing three times with PBS, samples were incubated with Cy5-conjugated goat anti-rabbit IgG or goat anti-mouse IgG (1:200; Abcam) in PBS for 1 h, and then stained with DAPI for 10 min at room temperature (Life Technologies). Fluorescence signals were imaged using a fluorescence microscope (Olympus IX71; Olympus).

Li G., Kang Y., Feng X., Wang G., Yuan Y., Li Z., Du L, & Xu B. (2023). Dynamic changes of enhancer and super enhancer landscape in degenerated nucleus pulposus cells. Life Science Alliance, 6(6), e202201854.

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Spinal Cord Protein Preparation and Analysis

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The proteins from the L4-5 segments of the spinal cord were prepared on the 3^rd day following injection as previously described (19 (link),20 (link)). Protein concentrations were determined using a Bicinchoninic Acid Assay Protein Assay kit (Thermo Fisher Scientific, Inc.). Proteins (30 µg) were separated by 8% SDS-PAGE gel and transferred onto a nitrocellulose membrane. The nitrocellulose membrane was blocked with 5% non-fat dry milk for 2 h at room temperature followed with a primary antibody against TLR4 (1:500; cat. no. ab13556; Abcam, Cambridge, MA, USA) at 4°C overnight, and then with secondary antibody (goat anti-mouse IgG; 1:1,000; cat. no. ab6789; Abcam) conjugated with horseradish peroxidase for 2 h at room temperature. The proteins were detected using Pierce ECL Plus Western Blotting Substrate (cat. no. 32134; Thermo Fisher Scientific, Inc.). β-actin (Sigma-Aldrich-Merck KGaA, Darmstadt, Germany; 1:500) was used as a loading control. Densitometry analysis was performed using ImageJ software (version 1.8.0; National Institutes of Health, Bethesda, MD, USA).

Liu Y., Zhang Y., Pan R., Chen M., Wang X., Kong E., Yu W., Sun Y, & Wu F. (2018). Lentiviral-mediated inducible silencing of TLR4 attenuates neuropathic pain in a rat model of chronic constriction injury. Molecular Medicine Reports, 18(6), 5545-5551.

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Immunoblotting Analysis of FLI1 Protein

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Proteins were extracted using standard procedures in the presence of protease inhibitors^{57 (link)}. Protein lysates were fractionated by SDS-PAGE and transferred to PVDF membranes (Millipore) using TransFi (ThermoFisher Scientific). Membranes were probed for FLI1 with a mouse monoclonal antibody (1/400; BD Pharmigen). GAPDH was used as a loading control (1/200; Abcam). Secondary antibodies were HRP-conjugated with a goat anti-mouse IgG (1/1000; Abcam) and blots were developed with the ECL reagent (GE Healthcare) and exposed to film (Kodak).

Martinez-Lage M., Torres-Ruiz R., Puig-Serra P., Moreno-Gaona P., Martin M.C., Moya F.J., Quintana-Bustamante O., Garcia-Silva S., Carcaboso A.M., Petazzi P., Bueno C., Mora J., Peinado H., Segovia J.C., Menendez P, & Rodriguez-Perales S. (2020). In vivo CRISPR/Cas9 targeting of fusion oncogenes for selective elimination of cancer cells. Nature Communications, 11, 5060.

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Immunohistochemical Analysis of Tumor Sections

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⁺Formalin-fixed tumor tissues were embedded in paraffin and sliced into 5 μm sections. The sections were stained with H&E and antibody. The sections were incubated with the mouse anti-Ki-67 and caspase-3 (1:500, Abcam) at room temperature for 1 h. Subsequently, the sections were washed 3 times in PBS and incubated for 30 min at room temperature with goat anti-mouse IgG (1:10000, Abcam). The sections were examined by light-microscopy at × 400 magnification.

Yin Q., Zhao N., Chang Y., Dong M., Xu M., Xu W., Jin H.F., Liu W, & Xu N. (2023). Melanoma stem cell vaccine induces effective tumor immunity against melanoma. Human Vaccines & Immunotherapeutics, 19(1), 2158670.

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Western Blot Analysis of OXPHOS Complexes

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Whole cells were lysed in buffer containing 1% SDS, 50 mM Tris– HCl (pH 7.8), 150 mM NaCl, and Complete Mini-Roche protease inhibitors. BCA Protein assay (Thermo Fisher) was used to determine and equalize total protein concentration across samples. Heat-denatured samples were loaded onto precast NuPAGE 4%–12% Bis-Tris gels (Thermo Fisher) at 15 μg of protein per well. Loaded gels were run for 1–2 h at 150 V to allow for size-dependent protein separation. Proteins were then transferred to a PVDF membrane and blocked in a 5% milk solution. Membranes were incubated with primary antibodies overnight at 4°C. Primary antibodies (OXPHOS Human WB Antibody Cocktail; Anti-TOMM20; Abcam) were diluted in 5% milk in PBS supplemented with 0.1% TWEEN20. Membranes were subsequently incubated with HRP-coupled secondary antibodies (Goat anti Rabbit IgG; Invitrogen; Goat Anti-Mouse IgG; Abcam). Proteins were detected using Amersham ECL Detection Reagent (Life Technologies) and imaged using Alliance chemiluminescence imaging system (UVITEC). Quantification of relative signal intensities of OXPHOS complexes were normalized to TOM20 levels.

Podmanicky O., Gao F., Munro B., Jennings M.J., Boczonadi V., Hathazi D., Mueller J.S, & Horvath R. (2023). Mitochondrial aminoacyl-tRNA synthetases trigger unique compensatory mechanisms in neurons. Human Molecular Genetics, 33(5), 435-447.

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Immunohistochemical Characterization of Cardiac Cells

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Immunohistochemical assessments were carried out on frozen cardiac tissue. Briefly, sections were fixed in 4.2 % paraformaldehyde (BD Biosciences, San Jose CA) for 15 mins, followed by permeabilization and blocking with normal goat serum for 30 minutes at room temperature. Slides were incubated with primary antibodies (Abcam, Cambridge, MA): rabbit anti-CD133 (Catalog # ab19898; used at 1:25), rat anti-CD45 (Catalog # ab30446; used at 1:25), and mouse anti-SSEA-4 (Catalog # ab16287; used at 1:20). The sections were then washed with PBS-Tween, and then incubated with all three secondary antibodies (Abcam, Cambridge, MA) at room temperature for 30 minutes: goat anti-rabbit IgG (Alexa Fluor 488; Catalog # ab150081; used at 1:200), goat anti-rat IgG (Alexa Fluor 647; Catalog # ab150167; used at 1:200), goat anti-mouse IgG (Alexa Fluor 555; Catalog # ab150118; used at 1:200). The sections were finally incubated with 0.1% Sudan Black B (Sigma Aldrich, St. Louis, MO) for 30 minutes. ~20 adjacent areas were imaged at 40x magnification using Nikon Confocal Microscope A1 (Nikon, Tokyo, Japan) in the University of Kentucky Confocal Microscopy facility. Cell numbers were expressed as cells/high power field (HPF). Cell numbers are expressed as cells/HPF.

El-Helw M., Chelvarajan L., Abo-Aly M., Soliman M., Milburn G., Conger A.L., Campbell K., Ratajczak M.Z, & Abdel-Latif A. (2020). Identification of Human Very Small Embryonic Like Stem Cells (VSELS) in human heart tissue among young and old individuals. Stem cell reviews and reports, 16(1), 181-185.

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Quantification and Analysis of Protein Signaling

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Total protein was drawn from mesenteric glands using a total protein extraction sample kit (Nanjing Keygen Biotech Co., Ltd. China). Protein quantification was performed using the BCA protein quantification kit (Nanjing Keygen Biotech Co., Ltd. China). The protein samples were resolved using SDS-PAGE. After electrophoresis, proteins were transferred and blocked in 5% skim milk (overnight at 4°C). The appropriate amounts of rat anti-Smad3, NFAT2, and Stat3 mAb (Abcam, USK) were added and blocked in the shaker for 2.5 hours at 37°C. The membranes were washed three times with 1×TBST. Goat anti-mouse IgG (Abcam, USK) was added. The membrane protein was posted on the X-ray film for exposure and developed in the developing machine. The signal intensity was analysed using grey-scale analysis software (ImageTool 3.0).

Wang K., Zhu T., Wang H., Yang J., Du S., Dong G., Pei Z, & Hu G. (2018). Adoptive Transfers of CD4+CD25+ Tregs Raise Foxp3 Expression and Alleviate Mouse Enteritis. BioMed Research International, 2018, 9064073.

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Quantitative Western Blot Analysis of Insulin and PDX1

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Sample cells were lysed using a RIPA protein extraction reagent (Beyotime, Beijing, China) supplemented with a protease inhibitor cocktail (Roche, Rotkreuz, Switzerland) and phenylmethylsulfonyl fluoride (Roche, Rotkreuz, Switzerland). A 50 μg sample of the extracted protein was separated via 15% SDS-PAGE, transferred to an Immobilon®-P Transfer membrane (Millipore, USA), and incubated with specific antibodies. The autoradiograms were quantified using densitometry (Quantity One software; Bio-Rad). After the protein content was determined, western blotting was performed. The specific antibodies and dilutions were as follows: anti-PDX1 (1 : 1000) and anti-INSULIN (1 : 800). All antibodies were obtained from Abcam (AB, USA). The target protein levels were normalized to anti-GAPDH (1 : 1000) from Abcam (AB, USA). The secondary antibodies used were horseradish peroxidase-conjugated goat anti-rabbit IgG (1 : 10,000) and goat anti-mouse IgG (1 : 5000) from Abcam (AB, USA).

Lu Z., Wang J., Wang X., Li Z., Niu D., Wang M., Xiang J., Yue Y., Xia Y, & Li X. (2021). miR-375 Promotes Pancreatic Differentiation In Vitro by Affecting Different Target Genes at Different Stages. Stem Cells International, 2021, 6642983.

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