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Anti mouse igg hrp conjugate

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The Anti-mouse IgG HRP conjugate is a reagent used in immunoassays and Western blotting applications. It consists of a horseradish peroxidase (HRP) enzyme conjugated to an antibody that specifically binds to mouse immunoglobulin G (IgG).

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Lab products found in correlation

Anti rabbit igg hrp conjugate, by Promega (10 mentions) Hybond p membrane, by GE Healthcare (3 mentions) Western blot ecl plus kit, by GE Healthcare (3 mentions) Trans blot turbo transfer system, by Bio-Rad (3 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions) Ripa buffer, by Merck Group (2 mentions) α tubulin, by Merck Group (2 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions) Anti goat igg hrp conjugate, by Promega (2 mentions) Anti rabbit immunoglobulins hrp, by Agilent Technologies (1 mentions) Rabbit anti sqstm1 p62, by Cell Signaling Technology (1 mentions) β actin mouse monoclonal antibody, by Santa Cruz Biotechnology (1 mentions) Restore western blot stripping buffer, by Thermo Fisher Scientific (1 mentions) Lps from salmonella typhosa, by Merck Group (1 mentions) Anti rabbit rhodamine conjugated tubulin, by Bio-Rad (1 mentions) Kta start protein purification system, by GE Healthcare (1 mentions) P chk1, by Cell Signaling Technology (1 mentions) Gapdh, by Promega (1 mentions) Dnmt3a, by Cell Signaling Technology (1 mentions) Stat1, by R&D Systems (1 mentions)

Close spelling variants of this product

Anti-rabbit and mouse IgG horseradish peroxidase (HRP) conjugates

25 protocols using anti mouse igg hrp conjugate

1

Western Blot Protein Detection

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After separation in SDS-PAGE, the proteins were transferred to the Hybond-P membrane (GE Healthcare, Chicago, IL, USA) by semi-dry transfer using the Trans-Blot Turbo Transfer System (Bio-Rad Laboratories, Hercules, CA, USA). To prevent non-specific binding of antibodies, the membrane was incubated in a 5% solution of skimmed dry milk in 10 mM PBS for 1 h with constant stirring. Then, the membrane was incubated with mouse anti-M2e polyclonal antibodies, washed, and then treated with the anti-mouse IgG HRP conjugate (W4021, Promega, Madison, WI, USA). Specific protein complexes were visualized using the Western Blot ECL Plus kit (GE Healthcare).
Zykova A.A., Blokhina E.A., Stepanova L.A., Shuklina M.A., Ozhereleva O.O., Tsybalova L.M., Kuprianov V.V, & Ravin N.V. (2023). Nanoparticles Carrying Conserved Regions of Influenza A Hemagglutinin, Nucleoprotein, and M2 Protein Elicit a Strong Humoral and T Cell Immune Response and Protect Animals from Infection. Molecules, 28(18), 6441.
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2

Western Blot Analysis of CXORF21 and p62

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Cell lysates were prepared in RIPA buffer (Sigma-Aldrich) and run on a SDS polyacrylamide gel for electrophoresis. Protein was transferred onto a nitrocellulose membrane and blocked in 5% milk-PBS solution. The rabbit polyclonal anti-CXORF21 antibody (Atlas Antibodies; HPA001185) was used at a concentration of 1:1000 and the secondary polyclonal swine anti-rabbit immunoglobulins/HRP (Dako; P0217) at 1:1000. Membranes were stripped by Restore™ Western Blot Stripping Buffer (Thermo Fisher) and re-probed with mouse monoclonal β-Actin antibody (Santa Cruz Biotechnology; sc-47778) at 1:4,000 and anti-mouse IgG HRP conjugate (Promega; W4028) at 1:5,000 or secondary goat anti-mouse IgG HRP conjugate (Invitrogen; A16078) at 1:10,000. ImageJ was used to calculate the density of the bands relative to the loading control. Rabbit anti-SQSTM1/p62 (Cell Signalling, 5114) was used at a concentration of 1:1000 and detected with secondary goat anti-rabbit IgG HRP conjugate (Invitrogen; A16110) at 1:10,000. Raw blots are presented in accompyning Source Data file.
Odhams C.A., Roberts A.L., Vester S.K., Duarte C.S., Beales C.T., Clarke A.J., Lindinger S., Daffern S.J., Zito A., Chen L., Jones L.L., Boteva L., Morris D.L., Small K.S., Fernando M.M., Cunninghame Graham D.S, & Vyse T.J. (2019). Interferon inducible X-linked gene CXorf21 may contribute to sexual dimorphism in Systemic Lupus Erythematosus. Nature Communications, 10, 2164.
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3

Complement Pathway Modulation by RaCI2

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The effect of RaCI2 on all three pathways was tested using a Wieslab complement system screen (Euro Diagnostica, Sweden) following manufacturer’s instructions. Measurements were subtracted with buffer only and normalised for serum only samples (100% activity). Other classical and alternative pathway assays were carried out as described by A. Roos et al. 42 (link), with the following modifications. 2 μg/ml IgM (Bio-Rad) and 1.8 mg/ml LPS from Salmonella typhosa (Sigma-Aldrich) were used to coat ELISA plates. Human serum was diluted 1/100 (classical pathway) or 1/10 (alternative pathway). MAC deposition was detected with primary antibodies against C5b-9 (aE11, Abcam), and secondary anti-mouse IgG HRP conjugate (Promega). HRP activity was detected with 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) liquid substrate (Sigma-Aldrich) and the reaction was stopped by adding an equal volume of 1% SDS. The absorption was measured at 405nm. Measurements were subtracted with buffer only samples and normalized for serum only samples (100% activity). The competition assays were performed using size-exclusion purified inhibited C5 complexes (see purification of C5 complexes above) added to the reaction at the concentrations specified.
Jore M.M., Johnson S., Sheppard D., Barber N.M., Li Y.I., Nunn M.A., Elmlund H, & Lea S.M. (2016). Structural basis for therapeutic inhibition of complement C5. Nature structural & molecular biology, 23(5), 378-386.
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4

CFTR Protein Expression and Detection

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Plasmids used for transient transfection expressed WT or mutant CFTR in the pcDNA5/FRT vector. Anti-CFTR antibodies used for detection were 217 and 596 (provided by J. Riordan, University of North Carolina, Chapel Hill, North Carolina; http://cftrantibodies.web.unc.edu/) each at 1:1000 and 1:500 working dilutions in immunoblotting buffer (5% bovine serum albumin [BSA] in Tris-buffered saline, pH 7.5, 0.1% Tween-20, and 0.1% NaN3) respectively. Antibodies used for immunoprecipitation was mAB 24–1 (ATCC HB-11947) purified from B lymphocyte hybridoma cells using a recombinant Protein G–Sepharose 4B Affinity Column on an ÄKTA start protein purification system (GE Life Science Product # 29022094). Secondary antibodies used were goat anti-mouse StarbrightB700 (Bio-Rad), anti-mouse IgG HRP conjugate (Promega) anti-rabbit rhodamine–conjugated tubulin (Bio-Rad).
Kim M., McDonald E.F., Sabusap C.M., Timalsina B., Joshi D., Hong J.S., Rab A., Sorscher E.J, & Plate L. (2023). Elexacaftor/VX-445-mediated CFTR interactome remodeling reveals differential correction driven by mutation-specific translational dynamics. bioRxiv.
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5

Ronin Knockout and UV-C Response Assays

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To detect Cre-mediated Ronin knockout, cells were plated at a density of 250,000 cells/20cm2 and fed for four days with ES cell medium supplemented with ethanol or 0.25 μM 4HT. To detect protein expression after UV-C treatment, cells were plated at a density of 1.1×106 cells/60cm2, fed as above for four days, and UV-C irradiated with 12 J/m2. Cells were harvested by trypsinization, washed with PBS and whole-cell protein extracts were prepared and Western blots performed as described previously (Dejosez et al., 2010 (link)). Antibodies used were: Ronin/Thap11 (BD Biosciences), α-Tubulin (Sigma), p-Chk1 (Ser345, Cell Signaling Technology), Chk1 (Cell Signaling Technology), p-p53 (hSer15/mSer18, Abcam), p53 (Leica Biosystems), Anti-Rabbit IgG HRP Conjugate (Promega), and Anti-Mouse IgG HRP Conjugate (Promega). Signal intensities were quantified with the Image J software.
Seifert B.A., Dejosez M, & Zwaka T.P. (2017). Ronin influences the DNA damage response in pluripotent stem cells. Stem cell research, 23, 98-104.
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6

Immunoblotting of DNMT3A and GAPDH

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Immunoblotting was performed as in ref12 (link). The following antibodies and dilutions were used: Primary antibodies: DNMT3A (Cell Signaling Technology, catalog no. 32578, D2H4B, 1:5,000), GAPDH (Santa Cruz Biotechnology, catalog no. sc-47724, 0411, 1:8,000). Secondary antibodies: anti-rabbit IgG HRP conjugate (Promega, catalog no. W4011, 1:100,000 for DNMT3A), anti-mouse IgG HRP conjugate (Promega, catalog no. W4021, 1:400,000 for GAPDH). Immunoblots were visualized using SuperSignal West Femto (DNMT3A) or SuperSignal West Pico PLUS (GAPDH) chemiluminescent substrates (Thermo Fisher Scientific).
Hampl V., Vaňáčová Š., Kulda J, & Flegr J. (2001). Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains. BMC Evolutionary Biology, 1, 11.
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7

Immunoblotting for STAT1, CDKs, and PARP

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Cells were treated with CA, ruxolitinib, or DMSO for 2 h. Pellets were collected and washed with PBS, then lysed with RIPA buffer (Sigma R0278) supplemented with protease inhibitors (Sigma P8340) and phosphatase inhibitors (Sigma P0044 and P5726). Proteins were resolved on NuPAGE 4–12% polyacrylamide gels in LDS buffer (Thermo Fisher), transferred to PVDF membranes with Tris-Glycine transfer buffer, blocked with 5% BSA or 5% milk in 0.1% TBST, then probed with antibodies. Primary antibodies included: STAT1 (CST #9172), STAT1 (R&D Systems #PAF-ST1), STAT1 pS727 (CST #9177), STAT1 pY701 (CST #9167), CDK8 (CST #4101 or #4106), CDK19 (Sigma #HPA007053), CDK9 (CST #2316), FLAG (Sigma F1804), PARP (CST #9532), actin (Sigma #A5060), GAPDH (Santa Cruz sc-47724). For secondary antibodies, we used either anti-rabbit IgG HRP conjugate (Promega #401B) or anti-mouse IgG HRP conjugate (Promega #402B).
Nitulescu I.I., Meyer S.C., Wen Q.J., Crispino J.D., Lemieux M.E., Levine R.L., Pelish H.E, & Shair M.D. (2017). Mediator Kinase Phosphorylation of STAT1 S727 Promotes Growth of Neoplasms With JAK-STAT Activation. EBioMedicine, 26, 112-125.
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8

Characterization of RNF183 in NF-κB Signaling

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The plasmid of pcDNA4-myc/his-RNF183 was constructed by the company of genewiz (Suzhou, China). A truncated form of RNF183 without amino acids 1–59 was generated by PCR and subsequent molecular cloning into pcDNA4-myc/his vector. IL-8 and IL-8-△NF-κB reported plasmids were kindly provided by Professor Hongbin Shu (Wuhan University, China). The antibodies used were listed as follows: human anti-RNF183 antibody (1:1000 for western blot, 1:200 for IHC, ab197321, Abcam, Cambridge, UK), anti-TAB1 antibody (1;1000, #3226 CST), anti-TRAF2 antibody (1:200, sc-136999, Santa Cruz Biotech, Santa Cruz, CA, USA), anti-TRAF6 (1:200, sc-7221, Santa Cruz), anti-NF-κB p65 antibody (1:1000 for western blot, 1:200 for IHC, 1:100 for CHIP, #8242, CST), anti-NF-κB P50 antibody (1:1000, #12540, CST, Boston, MA, USA), anti-ki-67 antibody(1:1000, #12202, CST), anti-β-actin (1:200, sc-47778, Santa Cruz), anti-mouse IgG HRP conjugate (1:8000, W402B, Promega, Madison, WI, USA), anti-Rabbit IgG HRP conjugate (1:2000, #7074P2, CST), normal rabbit IgG (1 μg for ChIP, sc-3888, Santa Cruz). Recombinant human IL-8 was purchased from Novus (Littleton, CO, USA) (NBP2-34905). The siRNAs (ribobio Company, Beijing, China) sequences were listed in supplementary table 1.
Geng R., Tan X., Wu J., Pan Z., Yi M., Shi W., Liu R., Yao C., Wang G., Lin J., Qiu L., Huang W, & Chen S. (2017). RNF183 promotes proliferation and metastasis of colorectal cancer cells via activation of NF-κB-IL-8 axis. Cell Death & Disease, 8(8), e2994-.
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9

Immunoblotting of Cellular Stress Markers

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Primary antibodies: Dicer (human), ATF4, CHOP, Caspase-3, total and phosphorylated eIF2α (Ser51), total and phosphorylated PKR (Thr451) (Cell Signaling Technology, Boston, MA); Dicer (mouse) (Bethyl laboratories, Montgomery, TX); GAPDH, HSP90, HSP70 (Abcam, Cambridge, MA). Secondary antibodies: Anti-mouse IgG HRP conjugate and anti-rabbit IgG HRP conjugates (Promega, Madison, WI).
Devasthanam A.S, & Tomasi T.B. (2017). Dicer protein levels elevated by mild hyperthermia promote a pro-survival phenotype. Oncotarget, 8(40), 67001-67016.
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10

Western Blot Analysis of Phosphorylated Proteins

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Protein extracts were prepared by lysing cells with RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS, 1mM Na3VO4, 1 mM NaF, 0.1 mM PMSF, 10 μM aprotinin, 5 μg/mL leupeptin, 1 μg/mL pepstatin A). Protein samples were diluted using 2x Laemmli loading buffer, mixed, and boiled for 5 minutes at 95°C. Samples were analyzed by SDS/PAGE using a 10% acrylamide gel, followed by transfer onto PVDF membranes (Millipore IPVH00010). Membranes were blocked with 5% BSA (ThermoFisher BP9706) in Tris-buffered saline (50 mM Tris-HCl pH 7.5, 150 mM NaCl) and 0.1% Tween-20 for 1 h at room temperature.
Primary antibody labeling was completed with anti-P-Akt (1:1,000; Cell Signaling 4060), anti-P-ERK (1:1000; Sigma M8159), anti-P-FOXO (1:1000; Millipore 07–695), or anti-actin (1:10,000; Sigma A2066) overnight at 4°C. Secondary antibody labeling was completed using anti-rabbit IgG-HRP conjugate (1:10,000; Promega W401B) or anti-mouse IgG-HRP conjugate (1:10,000; Promega W402B) by incubating membranes for 2 h at room temperature. Blots were imaged onto film using luminol enhancer (ThermoFisher 1862124). Densitometry analysis was completed using three independent blots using BioRad Image Lab and GraphPad Prism 9 with bands normalized to actin.
Trammell C.E., Ramirez G., Sanchez-Vargas I., St Clair L.A., Ratnayake O.C., Luckhart S., Perera R, & Goodman A.G. (2022). Coupled small molecules target RNA interference and JAK/STAT signaling to reduce Zika virus infection in Aedes aegypti. PLoS Pathogens, 18(4), e1010411.
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