Mab1259

Manufactured by Abcam

MAB1259 is a mouse monoclonal antibody designed for use as a research tool. It targets the human PCNA (Proliferating Cell Nuclear Antigen) protein. The antibody is suitable for applications such as immunocytochemistry and immunohistochemistry.

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Lab products found in correlation

Alexa dye, by Thermo Fisher Scientific (2 mentions) Operetta, by PerkinElmer (2 mentions) Ab97959, by Abcam (2 mentions) G3893, by Merck Group (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Ab45939, by Abcam (1 mentions) Cm3050 s cryostat, by Thermo Fisher Scientific (1 mentions) Superfrost plus slide, by Thermo Fisher Scientific (1 mentions) Ab32454, by Abcam (1 mentions) Ab9610, by R&D Systems (1 mentions) Ab13970, by Merck Group (1 mentions) Ab9610, by Abcam (1 mentions) Mab2018, by Santa Cruz Biotechnology (1 mentions)

3 protocols using mab1259

Quantifying Neural Precursor Cells

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Fixed cells were permeabilized with triton-x and, if applicable, treated according to the manufacturer's instructions to expose EdU staining. Primary antibodies (mouse monoclonal Nestin, R&D MAB1259, 1:500; rabbit SOX2, Abcam ab97959; 1:1000; rabbit vimentin, Abcam ab45939, 1:900), were added after a blocking step and incubated overnight at 4°C. Secondary antibodies, goat, conjugated to Alexa dyes (Molecular Probes), 1:1000, were left on for one hour at room temperature. Finally, cell nuclei were stained with DAPI (Sigma Aldrich). Plates were scanned with the Operetta (Perkin Elmer) and analyzed with the Harmony software. The number of cells was automatically counted with triplicates for each condition. Error bars in the graphs denote standard deviation. The effect of ALECSAT was estimated as the slope of the linear region of the dose response curve (logarithmic x-axis) for each treatment. P values are included where appropriate and P < .05 was considered significant. The Bonferroni correction was used to adjust for multiple testing. Pearson correlation (r) was calculated for correlations.

Wenger A., Werlenius K., Hallner A., Bergh Thorén F., Farahmand D., Tisell M., Smits A., Rydenhag B., Jakola A.S, & Carén H. (2017). Determinants for Effective ALECSAT Immunotherapy Treatment on Autologous Patient-Derived Glioblastoma Stem Cells. Neoplasia (New York, N.Y.), 20(1), 25-31.

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Immunostaining of Neural Stem Cell Markers

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Primary antibodies (GFAP monoclonal mouse antibody, Sigma-Aldrich G3893 1:1000; rabbit OLIG2, Chemicon AB9610, 1:300; mouse monoclonal nestin, R&D MAB1259, 1:500; rabbit SOX2, Abcam ab97959; 1:1000; rabbit MAP2, Abcam ab32454, 1:1000; rabbit vimentin, Abcam ab45939, 1:900), were incubated overnight at 4°C. Goat secondary antibodies conjugated to Alexa dyes (Molecular Probes), 1:1000, were added for 1 hr at room temperature and DAPI was used as a nuclear counterstain. For tumor spheres; 10-12 μm thick sections of embedded spheres were cut with a Leica CM3050S Cryostat and mounted on Superfrost™ Plus Slides (Fisher Scientific), which were stored in -20°C until staining and imaging as described above. The Operetta (Perkin Elmer) was used for acquisition of images and the Harmony software for quantification of cells.

Wenger A., Larsson S., Danielsson A., Elbæk K.J., Kettunen P., Tisell M., Sabel M., Lannering B., Nordborg C., Schepke E, & Carén H. (2017). Stem cell cultures derived from pediatric brain tumors accurately model the originating tumors. Oncotarget, 8(12), 18626-18639.

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Immunofluorescence Staining of Neural Cells

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Cells were fixed in 4% paraformaldehyde, permeabilised and blocked in 0.1% bovine serum albumin plus 3% goat serum solution. Samples were incubated overnight with primary antibodies followed by incubation with appropriate secondary antibodies (1:1000; Invitrogen) and 4′,6-diamidino-2-phenylindole (DAPI). Immunopositive cells were quantified using ∼5000 cells (minimum of ten random fields). Total cell number was determined by DAPI nuclear staining. The following primary antibodies were used: mouse nestin (1:10; Developmental Studies Hybridoma Bank, Rat-401), human nestin (1:500; R&D Systems, MAB1259), mouse Sox2 (1:100; Abcam, 92494), human Sox2 (1:50; R&D Systems, MAB2018), BLBP (also known as Fabp7) (1:200; Santa Cruz, sc-30088), GFAP (1:1000; Sigma, G3893), Tuj1 (also known as Tubb3) (1:1000; Biolegend, 801202), GFP (1:1000; Abcam, AB13970), Olig2 (1:400; Millipore, AB9610), mCherry (1:500; Abcam, AB167453), V5 tag (1:1000; eBioscience, 14679682), p53 (1:400; Cell Signaling, 2524).

Bressan R.B., Dewari P.S., Kalantzaki M., Gangoso E., Matjusaitis M., Garcia-Diaz C., Blin C., Grant V., Bulstrode H., Gogolok S., Skarnes W.C, & Pollard S.M. (2017). Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells. Development (Cambridge, England), 144(4), 635-648.

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