Mouse anti mcm2

Manufactured by BD

Mouse anti-MCM2 is a primary antibody that specifically binds to the MCM2 protein, a component of the MCM complex involved in the initiation of DNA replication. This antibody can be used for the detection and analysis of MCM2 in various applications, such as western blotting, immunohistochemistry, and immunocytochemistry.

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Anti-Mcm2 (3 citations) Mouse anti (2 citations)

3 protocols using mouse anti mcm2

Immunohistochemical Analysis of SVZ and Olfactory Bulb

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Brains were fixed overnight at 4°C in 4% PFA in PBS, then cryoprotected in 30% sucrose in PBS for 1–3 days at 4°C, then frozen in OCT or cryogel on dry ice after 3–12 hr equilibration at 4°C. Sections were cut at 12 μm thickness spanning the rostral half of the SVZ (typically 6 sections per slide) or the entire olfactory bulb (typically 8 sections per slide). Sections were immunostained with the following primary antibodies: rat anti-BrdU (Abcam clone Bu1/75, 1/500, after heat-mediated antigen retrieval), guinea pig anti-Dcx (1/1000; Millipore), mouse anti-Mcm2 (BD Biosciences, 1/500, after heat-mediated antigen retrieval), rat anti-Ki67 (1/500; eBioscience), mouse anti-tyrosine hydroxylase (1/1000; Millipore), rabbit anti-calretinin (1/1000; Sigma), rabbit anti-calbindin (1/500; Millipore), rabbit anti-S100β (1/1000; Dako), rabbit anti-GST-pi (1/3000; Enzo, Farmingdale, NY), and mouse anti-NeuN (1/1000; Millipore). Fixed whole-mount SVZs were stained with mouse anti-acetylated tubulin (1/1000; Sigma), rabbit anti-β-catenin (1/500; Sigma), mouse anti-GFAP (1/3000; Sigma), and goat anti-EGFR (1/250; R&D Systems). Alexa Fluor 488-, 555-, and 647-conjugated secondary antibodies were used (Life Technologies, Carlsbad, CA).

Mich J.K., Signer R.A., Nakada D., Pineda A., Burgess R.J., Vue T.Y., Johnson J.E, & Morrison S.J. (2014). Prospective identification of functionally distinct stem cells and neurosphere-initiating cells in adult mouse forebrain. eLife, 3, e02669.

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Immunohistochemical Quantification of Neurogenesis

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Tissue processing and immunohistochemistry was performed on free-floating sections according to standard published techniques (Villeda et al., 2014 (link)). Briefly, mice were anesthetized with 400 mg/kg chloral hydrate (Sigma-Aldrich) and transcardially perfused with 0.9% saline. Brains were removed and fixed in phosphate-buffered 4% paraformaldehyde (pH 7.4) at 4°C for 48 h before embedding in 30% sucrose for cryoprotection. Brains were then sectioned coronally at 40 Mm with a cryomicrotome (Leica Camera) and stored in cryoprotective medium. Sections were incubated overnight with mouse anti-MCM2 (1:500; 610700; clone: 46/BM28; BD Biosciences) or goat anti-DCX (1:7,500; Santa Cruz Biotechnology) primary antibodies, and revealed using appropriate A488-conjugated fluorescent secondary antibody (Life Technologies). To estimate the total number of positive cells per DG, immunopositive cells in the granule cell and subgranular cell layer of the DG were counted in every sixth coronal hemibrain section through the hippocampus for a total of six sections and multiplied by 12.

Ho T.T., Dellorusso P.V., Verovskaya E.V., Bakker S.T., Flach J., Smith L.K., Ventura P.B., Lansinger O.M., Hérault A., Zhang S.Y., Kang Y.A., Mitchell C.A., Villeda S.A, & Passegué E. (2021). Aged hematopoietic stem cells are refractory to bloodborne systemic rejuvenation interventions. The Journal of Experimental Medicine, 218(7), e20210223.

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Immunohistochemistry for Protein Localization

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Immunohistochemistry was performed as previously described^{54 (link)}. 3,3-Diaminobenzidine (Sigma) stains were performed using an ABC labeling kit (Vector Laboratories, Burlingame, CA). Primary antibodies were chosen according to previous studies in our lab or reports in the literature or instructions from the vendors: Rabbit anti-P-Smad2 (1:1000; Chemicon, AB3849), rabbit anti-eGFP (1:500; Life Technologies, A11122), chicken anti-eGFP (1:1000, Avis, GFP-1020), mouse anti-mCherry (1:200, Clontech, 632543), chicken anti-Tbr2 (1:500, Millipore, AB15894), mouse anti-MCM2 (1:500, BD Biosciences, 610700), rat anti-BrdU (1:1000; Abcam, AB6326), goat anti-DCX (1:500; Santa Cruz Biotechnology, sc-8066), mouse anti-PCNA (1:200; DAKO, M0879), rabbit anti-GFAP (1:1000; DAKO, Z0334), mouse anti-NeuN (1:1000; Millipore, MAB377), goat anti-Sox2 (1:200, Santa Cluz Biotechnology, sc-17320), mouse anti-HA (1:1000; Covance, MMS-101P), rabbit anti-c-fos (1:10000; Millipore, PC38). Antigen retrieval with 3 mol/L HCl was used for BrdU. For fluorescent stains, secondary antibodies were purchased from either Molecular Probes or Jackson Immunoresearch. For fluorescent staining procedures, final washes included the nuclear stain Topro-3 (1:500; Molecular Probes) or DAPI (5 μg/mL, Sigma) for 0.5 h at room temperature.

He Y., Zhang H., Yung A., Villeda S.A., Jaeger P.A., Olayiwola O., Fainberg N, & Wyss-Coray T. (2014). ALK5-dependent TGF-β signaling is a major determinant of late stage adult neurogenesis. Nature neuroscience, 17(7), 943-952.

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