Hrp linked anti mouse igg

Manufactured by GE Healthcare

Sourced in United States

The HRP-linked anti-mouse IgG is a laboratory reagent used in immunoassays. It consists of an anti-mouse immunoglobulin G (IgG) antibody conjugated to horseradish peroxidase (HRP), an enzyme that can be used to detect and quantify the presence of mouse IgG in samples.

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Lab products found in correlation

Chemidoc imaging system, by Bio-Rad (3 mentions) Hrp linked anti rabbit igg, by GE Healthcare (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) β actin, by Merck Group (2 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Anti cleaved parp antibody, by Cell Signaling Technology (1 mentions) Anti actb antibody, by Merck Group (1 mentions) Anti p21waf1 cip1 antibody, by Cell Signaling Technology (1 mentions) Anti bax antibody, by Cell Signaling Technology (1 mentions) Ecl prime western blotting detection kit, by GE Healthcare (1 mentions) Immun blot pvdf membrane, by Bio-Rad (1 mentions) Sds page, by Bio-Rad (1 mentions) Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions) Ecl plus western blot detection reagent, by GE Healthcare (1 mentions) Fastdigest restriction enzyme, by Thermo Fisher Scientific (1 mentions) Pbad myc his a plasmid, by Thermo Fisher Scientific (1 mentions) Ni nta agarose, by Qiagen (1 mentions) Cs 56, by Merck Group (1 mentions)

Close spelling variants of this product

HRP anti-mouse

17 protocols using hrp linked anti mouse igg

Immunoblotting Analysis of Apoptosis Regulators

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Total proteins were extracted from cultured cells using RIPA buffer containing protease inhibitor and phosphatase inhibitor (Thermo Fisher Scientific, Waltham, MA, USA). Proteins were subjected to 12% SDS‐PAGE (Bio‐Rad, Hercules, CA, USA). The separated proteins were transferred to Immun‐Blot PVDF membranes (Bio‐Rad) and incubated with anti‐ACTB antibody (A2066; Sigma‐Aldrich; 1:1000 dilution), anti‐cleaved PARP antibody (#5625; Cell Signaling Technology, Beverly, MA, USA; 1:1000 dilution), anti‐p21Waf1/Cip1 antibody (#2947; Cell Signaling Technology; 1:1000 dilution), anti‐Bax antibody (#5023; Cell Signaling Technology, Beverly, MA, USA; 1:1000 dilution), anti‐p53 antibody (Clone DO‐7; DAKO Cytomation, Carpinteria, CA, USA; 1:1000 dilution) and anti‐Noxa antibody (OP180; Calbiochem, Darmstadt, Germany; 1:1000 dilution) at 4°C overnight. Next, the membranes were incubated with HRP‐linked anti‐rabbit IgG (GE Healthcare Biosciences, Piscataway, NJ, USA) and HRP‐linked anti‐mouse IgG (GE Healthcare Biosciences) at a dilution of 1:100 000 for 1 hour at room temperature. The antigen–antibody complex was detected using an ECL Prime Western Blotting Detection Kit (GE Healthcare Biosciences).

Furukawa H., Makino T., Yamasaki M., Tanaka K., Miyazaki Y., Takahashi T., Kurokawa Y., Nakajima K., Takiguchi S., Mori M, & Doki Y. (2017). PRIMA‐1 induces p53‐mediated apoptosis by upregulating Noxa in esophageal squamous cell carcinoma with TP53 missense mutation. Cancer Science, 109(2), 412-421.

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Recombinant Protein Expression in E. coli

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E. coli O86:K61:B7 (ATCC 12701) was obtained from American Type Culture Collection (Rockville, MD, USA). E. coli DH5 [lacZΔM15 hsdR recA] competent cells were obtained from Gibco-BRL Life Technology. E. coli C43 (DE3) competent cells were obtained from Lucigen Co. (Middleton, WI). Nitrocellulose membrane, PCR reagents, and pBAD/Myc-His A plasmid were obtained from Invitrogen (Carlsbad, CA). Detergents were purchased from Anatrace (Maumee, OH). Ni-NTA agarose was from Qiagen (Valencia, CA). Anti-E. coli O86 antiserum (from Rabbit) was from Tianjin Biochip Corporation (Tianjin, China). HRP-linked anti-mouse IgG, anti-rabbit IgG, and ECL plus Western Blot detection reagents were purchased from GE healthcare (Piscataway, NJ). FastDigest restriction enzymes were purchased from Fermentas (Glen Burnie, MD). All other chemicals and solvents were purchased from Sigma-Aldrich.

Zhao G., Wu B., Li L, & Wang P.G. (2014). O-antigen Polymerase Adopts a Distributive Mechanism for Lipopolysaccharide Biosynthesis. Applied microbiology and biotechnology, 98(9), 4075-4081.

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Western Blot Analysis of Smad Signaling

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Protein lysates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred to a PVDF membrane. Membranes were blocked and incubated, according to the instructions provided, with the primary antibody. Protein bands were identified through a horseradish peroxidase-conjugated secondary antibody and enhanced chemiluminescence reagents (Pierce, Rockford, IL). The following primary antibodies were used for Western blots: phospho-Smad2 (Ser465/467) #3108, Smad2 #5339, phospho-Smad3 (Ser423/425) #9520, and Smad3 #9523 (each 1:1000, Cell Signaling Technology); CS-56 (1:500, C8035, Sigma-Aldrich); β-actin (1:3000, A5316, Sigma-Aldrich); GFAP (1:1000, Z0334, Dako); KCa3.1 (1:500, ab83740, Abcam). The following secondary antibodies were used: HRP-linked anti-rabbit IgG (1:2000, NA9340V, GE Healthcare); HRP-linked anti-mouse IgG (1:2000, NA9310V, GE Healthcare). Immunoreactivity for each protein was quantified using Image J software. Blots of cell lysate were reprobed with β-actin as a loading control.

Yu Z., Yu P., Chen H, & Geller H.M. (2014). Targeted inhibition of KCa3.1 attenuates TGF-β-induced reactive astrogliosis through the Smad2/3 signaling pathway. Journal of neurochemistry, 130(1), 41-49.

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Western Blot Analysis of Reelin and Lipoprotein Receptors

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Protein samples from plasma, tissues or HAEC whole cell lysates were prepared in RIPA buffer and separated by SDS-PAGE. After transfer onto nitrocellulose membranes (Bio-Rad), blots were probed separately with antibodies against mouse Reelin (G10), mouse Apoer2 and Vldlr as indicated. The G10 antibody was a generous gift from Andre Goffinet (Université catholique de Louvain, Belgium) (54 (link)). The function blocking CR50 antibody was kindly provided by Dr. Katsuhiko Mikoshiba (RIKEN, Saitama, Japan). The Vldlr antibody was purchased from Millipore Corporation (Cat #MABS25) and the Apoer2 antibody was generated in our laboratory as described previously (55 (link)). Antibodies against IκBα (Cat #4812) and β-actin (Cat #8457) were purchased from Cell Signaling Technology. Antibody against phospho-Ser³²/Ser³⁶-IκBα (Cat #ab12135) was purchased from Abcam. The secondary antibody used was HRP-linked anti-mouse IgG or anti-rabbit IgG (GE Healthcare), and membranes were visualized with SuperSignal West Pico Chemiluminescence reagents and X-ray film. Band intensity was quantified using scanning densitometry of non-saturating autoradiograms with ImageJ software (NIH) within linear exposure range.

Ding Y., Huang L., Xian X., Yuhanna I.S., Wasser C.R., Frotscher M., Mineo C., Shaul P.W, & Herz J. (2016). Loss of Reelin protects against atherosclerosis by reducing leukocyte-endothelial adhesion and lesion macrophage accumulation. Science signaling, 9(419), ra29.

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Western Blot Analysis of CaMKII, PSD-95, and GluN2B

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Before undergoing SDS-PAGE, samples were boiled in Laemmli sample buffer for 5 min at 95° C. Proteins were separated in a resolving phase polymerized from 10% acrylamide, then transferred to a polyvinylidene difluoride membrane at 24 V for 1 hour at 4° C. Membranes were blocked in 5% milk and incubated with mouse anti-CaMKII (1:2000; BD Biosciences, RRID: AB_398819), mouse anti PSD-95 (1:2500; NeuroMab, RRID: AB_10698024) or mouse anti-GluN2B (1:1000; Cell Signaling Technology, RRID: AB_2798506) followed by incubation with HRP-linked anti-mouse IgG (1:6000; GE Healthcare, RRID: AB_772210). Blots were developed using chemiluminescence (Super Signal West Femto, Thermo-Fisher), imaged using the Chemi-Imager 4400 system (Alpha-Innotech), and analyzed by densitometry (Alphaese FC).

Batchelor M., Knutton S., Caprioli A., Huter V., Zanial M., Dougan G, & Frankel G. (1999). Development of a Universal Intimin Antiserum and PCR Primers. Journal of Clinical Microbiology, 37(12), 3822-3827.

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Western Blot Analysis of β-Catenin

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Cells (200,000 per sample) were lysed in Laemmli sample buffer (Bio-Rad Laboratories, Hercules, CA, USA), heated for 5 min at 95 °C, separated on Mini Protean TGX precast gels (Bio-Rad), and thereafter electroblotted onto nitrocellulose membranes (Bio-Rad). The membranes were blocked for 1 h with 5% BSA in PBS with 0.1% Tween-20. β-Catenin was detected using a β-catenin antibody (clone 14/Beta-Catenin, BD Biosciences) followed by HRP-linked anti-mouse IgG (GE Healthcare, Uppsala, Sweden). The Western blots were visualized by chemiluminescence (GE Healthcare) on a ChemiDoc Imaging System (Bio-Rad). The membranes were stripped using Re-Blot Plus Mild (Millipore) followed by reblocking and detection of β-Actin using an HRP-conjugated β-Actin antibody (clone C4, Santa Cruz Biotechnology, Dallas, TX, USA).

Tebroke J., Lieverse J.E., Säfholm J., Schulte G., Nilsson G, & Rönnberg E. (2019). Wnt-3a Induces Cytokine Release in Human Mast Cells. Cells, 8(11), 1372.

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Viral Plaque Assay for X47 Influenza

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Complete lungs were harvested on day 6 after infection and homogenized in 1 mL of PBS using a sterile metal bead on the Mixer Mill MM 200 (Retsch). Next, the lung homogenates were cleared by centrifuging at 1,000 × g for 10 min at 4°C. A serial dilution series of the cleared lung homogenates, made in serum-free DMEM medium, was added to a monolayer of Madin-Darby Canine Kidney (MDCK) cells in a 6-well plate (1 million cells per well). 1 h later, the cells were overlaid with an equal volume of 1.2% Avicel RC-591 (FMC Biopolymer) supplemented with 4 μg/mL of L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (Sigma). Plates were incubated at 37°C in 5% CO₂ for 3 days, after which the overlay was removed and cells were fixed with 4% paraformaldehyde. Viral plaques were stained using convalescent mouse anti-X47 serum followed by HRP-linked anti-mouse IgG (NXA931, GE Healthcare). After washing, the plaques were visualized with TrueBlue peroxidase substrate and counted (KPL, Gaithersburg, MD, USA).

Van Hoecke L., Verbeke R., De Vlieger D., Dewitte H., Roose K., Van Nevel S., Krysko O., Bachert C., Schepens B., Lentacker I, & Saelens X. (2020). mRNA Encoding a Bispecific Single Domain Antibody Construct Protects against Influenza A Virus Infection in Mice. Molecular Therapy. Nucleic Acids, 20, 777-787.

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Therapeutic Agents and Antibody Protocols

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17-(Allylamino)-17-demethoxygeldanamycin (17-AAG), the NQO1 inhibitor ES936, cis-diammineplatinum (II) dichloride (CDDP), and dacarbazine (DTIC) were obtained from Sigma Aldrich. 17-AAG and ES936 were prepared at a stock concentration of 10 mM in dimethyl sulfoxide. CDDP was directly dissolved in culture medium. DTIC was prepared as a 0.1 M stock in 0.1 M HCl.
The antibody against NRF2 was obtained from Abcam (ab-62352, Cambridge, MA). The antibody against NQO1 was from Cell Signaling Technology (#3187, Boston, MA). α-tubulin antibody was from Sigma Aldrich (T5168). GAPDH antibody was from Covance (MMS-580S, Princeton, NJ). HRP-linked anti-mouse IgG and anti-rabbit IgG were from GE Healthcare (Waukesha, WI).

Kasai S., Arakawa N., Okubo A., Shigeeda W., Yasuhira S., Masuda T., Akasaka T., Shibazaki M, & Maesawa C. (2016). NAD(P)H:Quinone Oxidoreductase-1 Expression Sensitizes Malignant Melanoma Cells to the HSP90 Inhibitor 17-AAG. PLoS ONE, 11(4), e0153181.

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Western Blot Analysis of Tax Protein

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Cells were lysed with sample buffer (10 mM HEPES, pH 7.9, 500 mM NaCl, 3 mM MgCl2, 1 mM DTT, 1 mM PMSF, and 0.5% Triton X-100 supplemented with protease inhibitors). After incubation on ice for 60 min, whole cell lysates were centrifuged at 15,000 g for 10 min at 4°C to remove the debris. Protein concentration of the cleared lysates was determined using the Bradford assay. Cell lysates (15μg) were size-separated by electrophoresis on a 12% SDS-polyacrylamide gel (3h migration at 20 mA) and transferred onto PVDF membranes. The blot was blocked in PBS-5% milk and incubated with anti-Tax antibodies (1:1,000; kind gift of B. Cullen), anti-β-actin (1:5,000) obtained from Sigma-Aldrich (clone AC-15). After several washes, horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (1:10,000; Cell Signaling, The Netherlands) or HRP-linked anti-mouse IgG (1:10,000; GE Healthcare, France) were added to the membranes which were washed again several times and subsequently incubated with the Western Lightning ECL solution (Thermo Fisher Scientific, France). Images were captured using a ChemiDoc Imaging system (Biorad, France).

Pérès E., Blin J., Ricci E.P., Artesi M., Hahaut V., Van den Broeke A., Corbin A., Gazzolo L., Ratner L., Jalinot P, & Duc Dodon M. (2018). PDZ domain-binding motif of Tax sustains T-cell proliferation in HTLV-1-infected humanized mice. PLoS Pathogens, 14(3), e1006933.

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Mitochondrial Protein Analysis in Mouse Tissues

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The mice were heparinized (200 U/mouse, i.p.) and sacrificed by cervical dislocation. The liver, kidney and left ventricle were quickly excised after thoracotomy. Mitochondria were isolated by a differential centrifugation method and Western blot analyses were performed as described previously [25 (link),27 (link)]. The primary antibodies used were: custom-made anti-NCLX (1:500, [25 (link)]), anti-MCU (1:1000, Cell Signaling Technology, Danvers, MA, USA, #14997), and anti-COX IV (1:2000, Abcam, Cambridge, UK, ab14744). The secondary antibodies used were: HRP-linked anti-rabbit IgG (Jackson ImmunoResearch, West Grove, PA, USA) for the NCLX and MCU, and HRP-linked anti-mouse IgG (GE healthcare, Chicago, IL, USA) for the COX IV. The quality of the custom-made anti-NCLX antibody, which was verified using HeLa cells transiently expressing NCLX and NCLX–FLAG [25 (link)], was further confirmed in Western blot analysis using cardiac mitochondria, where a single positive band around 65 kDa was diminished by pre-incubating the antibody with an excess amount of antigen peptide (Figure S1).

Takeuchi A, & Matsuoka S. (2022). Spatial and Functional Crosstalk between the Mitochondrial Na+-Ca2+ Exchanger NCLX and the Sarcoplasmic Reticulum Ca2+ Pump SERCA in Cardiomyocytes. International Journal of Molecular Sciences, 23(14), 7948.

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