Normal mouse igg

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

Normal mouse IgG is a laboratory reagent used as a control in various immunological assays. It serves as a non-specific antibody control, representing the general immunoglobulin profile of a normal mouse. This product is intended for research use only and its specific applications should be determined by the end-user.

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Lab products found in correlation

227 protocols using normal mouse igg

Chromatin Immunoprecipitation Protocol

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One-mL chromatin-protein aliquots were precleared with 100 μL 50% DNA/Protein A/G slurry (Salmon Sperm DNA/Protein G Agar, catalog # 16-201; SSDNA/Protein A Agar, catalog # 16-157; Normal Mouse IGG, catalog # 12-371, EMD-Millipore Corporation, Billerica, MA). Precleared lysates were incubated overnight at 4 °C with rotation with 4 μg of anti-H3AcK9 (Anti-Acetyl Histidine H3-lys 9, catalog # 06-942 from Millipore) or 4 μg of Normal Mouse IGG (Normal Mouse IGG, catalog #12-371). Immunocomplexes were transferred to 1.5-mL tubes containing 100 μL of Salmon Sperm DNA/Protein A/G Agarose-50% slurry and incubated with rotation at 4 °C for 4 h. Agarose-immunocomplexes were pelleted by centrifugation at 750 rpm for 1 min at 4 °C. The supernatant was carefully removed and the agarose-immunocomplexes were subjected to two washes with low-salt buffer (150 mM NaCl, pH 8.0), two washes with high-salt buffer (500 mM NaCl, pH 8.0) and two washes with LiCl buffer (250 mM LiCl, pH 8.0), followed by one wash with Tris-EDTA (TE).

Restrepo R.J., Lim R.W., Korthuis R.J, & Shukla S.D. (2017). Binge alcohol alters PNPLA3 levels in liver through epigenetic mechanism involving histone H3 acetylation. Alcohol (Fayetteville, N.Y.), 60, 77-82.

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Validation of SNHG16-miR-488 Interaction

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Correlation between SNHG16 and miR-488 was verified via EZ-Magna RIP RNA-binding protein immunoprecipitation kit (Millipore) following the procedure of manufacturers. In brief, osteosarcoma cells transfected with miR-488 mimics were lysed with RIP lysis buffer, and 200 μL cell lysate was subjected to the incubation with RIP immunoprecipitation buffer containing magnetic beads that conjugated with human anti-Argonaute2 (Ago2) antibody (Millipore). Normal mouse IgG (Millipore) was applied as negative control. The enrichment of SNHG16 and PVT1 (positive control) was tested through qRT-PCR.

Bu J., Guo R., Xu X.Z., Luo Y, & Liu J.F. (2021). LncRNA SNHG16 promotes epithelial-mesenchymal transition by upregulating ITGA6 through miR-488 inhibition in osteosarcoma. Journal of Bone Oncology, 27, 100348.

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RNA Immunoprecipitation Assay Protocol

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EZ-Magna RIP kit (Millipore, Billerica, MA) was applied to conduct RIP assay according to the manufacturer’s protocol. Whole cell lysate were incubated with RIP magnetic beads conjugated with anti-Argonaute2 antibody and normal mouse IgG (Millipore). After incubating with Proteinase K buffer, immunoprecipitated RNA was obtained. Finally, the presence of the binding targets was validated by qRT-PCR.

Song Y., Shao L., Xue Y., Ruan X., Liu X., Yang C., Zheng J., Shen S., Chen J., Li Z, & Liu Y. (2019). Inhibition of the aberrant A1CF-FAM224A-miR-590-3p-ZNF143 positive feedback loop attenuated malignant biological behaviors of glioma cells. Journal of Experimental & Clinical Cancer Research : CR, 38, 248.

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Immunoprecipitation and Argonaute2 Interaction Profiling

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Cells were lysed in complete RNA lysis buffer containing magnetic beads conjugated with human anti-MOV10, anti-MOV10 antibody or negative control normal mouse IgG. And whole cell lysate of the control groups and anti-miR-103a-3p/anti-miR-382-5p groups were incubated with RIP immunoprecipitation buffer containing magnetic beads conjugated with human anti-Argonaute2 (Ago2) antibody (Millipore), and NC normal mouse IgG (Millipore). The samples were incubated with Proteinase K and then immunoprecipitated RNA was isolated. Purified RNA was obtained and then applied to quantitative PCR with reverse transcription analysis.

He Q., Zhao L., Liu X., Zheng J., Liu Y., Liu L., Ma J., Cai H., Li Z, & Xue Y. (2019). MOV10 binding circ-DICER1 regulates the angiogenesis of glioma via miR-103a-3p/miR-382-5p mediated ZIC4 expression change. Journal of Experimental & Clinical Cancer Research : CR, 38, 9.

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Immunoprecipitation of Argonaute Proteins

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The fat body of the B. bassiana ARSEF252 infected mosquitoes at 36 , 60, and 84 hpi were dissected, mixed, and homogenized in ice-cold RIP lysis buffer^{39 (link)}. Uninfected mosquitoes mixed with B. bassiana ARSEF252 mycelia were used as a control. Magnetic beads were pre-incubated with 5 μg of custom-made antibody against A. stephensi AGO1 (GenScript) (Supplementary Fig. 12) or normal mouse IgG (Millipore). Then, the antibody-coupled magnetic beads were incubated with 100 μl mosquito homogenates. The immunoprecipitates were pulled down and digested with protease K to release the bound sRNAs. Finally, the cDNAs were reverse transcribed using miRcute miRNA First-Strand cDNA Synthesis Kit (Tiangen). RT-PCR reactions were performed using a miRcute miRNA qPCR detection kit (Tiangen).

Cui C., Wang Y., Liu J., Zhao J., Sun P, & Wang S. (2019). A fungal pathogen deploys a small silencing RNA that attenuates mosquito immunity and facilitates infection. Nature Communications, 10, 4298.

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Hydroxymethylated DNA Immunoprecipitation Protocol

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hMeDIP was performed according to the manual's protocol (Diagenode). The antibodies used are as follows: 5hmC antibody (633HMC-100, Diagenode), Normal mouse IgG (12-371, Millipore). The primers for MeDIP/hMeDIP were presented in Supplemental Table S5.

Jiang Y., Yan B., Lai W., Shi Y., Xiao D., Jia J., Liu S., Li H., Lu J., Li Z., Chen L., Chen X., Sun L., Muegge K., Cao Y, & Tao Y. (2015). Repression of Hox genes by LMP1 in nasopharyngeal carcinoma and modulation of glycolytic pathway genes by HoxC8. Oncogene, 34(50), 6079-6091.

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Chromatin Immunoprecipitation of HPV-31

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Chromatin immunoprecipitations were performed using CIN612 or stable HPV-31 (WT or L2 Mutant 3X) cells grown as described above. Before fixation, cells were treated with versene to remove J2 fibroblast feeders. Cells were then seeded and grown to 80% confluency. Chromatin-immunoprecipitation was performed as previously described using SMC1 (Abcam, Cat. #ab9262), γ-H2AX (Millipore Cat. #05–636), CTCF (Millipore, Cat. #07–729), Normal Rabbit IgG (Santa Cruz, Cat. #SC-2027), Normal Mouse IgG Antibody (Santa Cruz, Cat. #SC-2025), Normal Mouse IgG (Millipore, Cat. #12-371B) and Protein-G Dynabeads (Invitrogen Cat. #100-03D) [50 (link)]. Real-time, touchdown PCR was performed with the Lightcycler 480 (Roche) and the HPV-31 CTCF Trio Primers and URR Primers; Forward: 5’-TTTGGTGGGTTGGGTATTGG-3’, Reverse:5’-GTAGGAGGCTGCAATACAGATG-3’. Forward, 5’-AACTGCCAAGGTTGTGTCATGC 3’, Reverse, 5’-TGGCGTCTGTAGGTTTGCAC-3’.

Mehta K., Gunasekharan V., Satsuka A, & Laimins L.A. (2015). Human Papillomaviruses Activate and Recruit SMC1 Cohesin Proteins for the Differentiation-Dependent Life Cycle through Association with CTCF Insulators. PLoS Pathogens, 11(4), e1004763.

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RIP Assay for RNA-Binding Proteins

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RIP assays were performed using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore) according to the manufacturer’s protocol. Normal mouse IgG (Millipore) was used as a negative control. Immunoprecipitated RNAs were reverse transcribed using SuperScript III First-Strand Synthesis System for RT-PCR (Invitrogen) according to the manufacturer’s instructions. The amount of each mRNA was quantified using the TaqMan assay.

Yugami M., Okano H., Nakanishi A, & Yano M. (2020). Analysis of the nucleocytoplasmic shuttling RNA-binding protein HNRNPU using optimized HITS-CLIP method. PLoS ONE, 15(4), e0231450.

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ChIP-PCR Analysis of ZEB2 Binding

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Protein-DNA complexes were immunoprecipitated from U251 and U87 cells using a Chromatin Immunoprecipitation Kit (Millipore, Billerica, MA, USA) according to the manufacturer’s protocol with anti-ZEB2 (Bethyl Laboratories, Montgomery, TX, USA) and normal mouse IgG (Millipore) polyclonal antibodies, the latter serving as a control for nonspecific DNA binding. The precipitated DNA was then subjected to PCR with specific primers to amplify the putative ZEB2 binding region analyzed via electrophoresis (Table S1).

Zeng Y., Que T., Lin J., Zhan Z., Xu A., Wu Z., Xie C., Luo J., Ding S., Long H., Zhang X, & Song Y. (2021). Oncogenic ZEB2/miR-637/HMGA1 signaling axis targeting vimentin promotes the malignant phenotype of glioma. Molecular Therapy. Nucleic Acids, 23, 769-782.

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Chromatin Immunoprecipitation (ChIP) Assay

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The following Abs were used for ChIP: α-macroH2A1.2 (Millipore MABE61), α-macroH2A1 (Millipore 07–219), α-phospho-H2AX (Millipore 05–636), α-H2AX (Abcam ab20669), α-H2AZ (Abcam ab4174), α-H2B (Abcam ab52484), normal mouse IgG (Millipore 12–371). Antibodies for IP were α-Flag M2 (Sigma F1804) and α-BRCA1 (Santa Cruz sc-6954). Primary antibodies for western blotting were: α-macroH2A1.2 (Millipore MABE61), α-phospho-H2AX (S139) (Millipore 05–636), α-BRCA1 (Santa Cruz sc-6954), α-PCNA (Santa Cruz sc-56), α-GAPDH (Santa Cruz sc-32233), α-SPT16 (Santa Cruz sc-28734), α-EZH2 (CST 5246s), α-SSRP1 (CST 134215), α-phospho-ATM (CST 4526s), α-phospho-CHK2 (CST 2661s), α-phospho-RPA S4/8 (Bethyl A300-245A), α-RPA2 (Millipore NA19L), α-H2AX (Abcam ab20669), α-H2AZ (Abcam ab4174), α-H2A (CST 12349s, Abcam ab15653). Secondary antibodies were goat anti-mouse IgG-HRP (Santa Cruz sc-2005, Invitrogen 31430) and goat anti-rabbit IgG-HRP (Santa Cruz sc-2004, Invitrogen 31460, CST 7074). Primary antibodies for IF were: α-γ-H2AX (Abcam ab11174, Milipore 05–636), α-BRCA1 (Santa Cruz, sc-6954), α-macroH2A1.2 (Millipore, MABE61), α-pRPA S33 (Bethyl, A300-246A), α-FANCD2 (Novus NB100-182). Secondary antibodies were from Life Technologies (goat-α-mouse or goat-α-rabbit conjugated to Alexa Fluor 488, 568, or 647).

Kim J., Sturgill D., Sebastian R., Khurana S., Tran A.D., Edwards G.B., Kruswick A., Burkett S., Hosogane E.K., Hannon W.W., Weyemi U., Bonner W.M., Luger K, & Oberdoerffer P. (2017). Replication stress shapes a protective chromatin environment across fragile genomic regions. Molecular cell, 69(1), 36-47.e7.

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