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Ab23345

Manufactured by R&D Systems
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Sourced in Japan

Ab23345 is a laboratory equipment product offered by R&D Systems. It is a device designed for specific functions within a research or testing environment. The core function of this product is to facilitate certain laboratory procedures, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Prb 278p, by Merck Group (1 mentions) Anti elf5, by Santa Cruz Biotechnology (1 mentions) Mab6166, by R&D Systems (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Af3797, by R&D Systems (1 mentions) Magna rip kit, by Merck Group (1 mentions) Af1924, by Abcam (1 mentions)

3 protocols using ab23345

1

Immunohistochemical Profiling of Neural Markers

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Immunohistochemistry was performed as described previously with slight modifications39 (link)43 (link)44 (link). Coronal sections (50 μm) were made using a cryostat, permeabilized with 0.1–0.5% Triton X-100/PBS, and incubated overnight with primary antibodies, which included anti-Tbr2 antibody (Abcam, ab23345), anti-Sox2 antibody (R&D Systems, AF2018), anti-Pax6 antibody (Covance, PRB-278P), anti-NeuN antibody (Chemicon, MAB377), anti-FOXP2 antibody (Atlas, HPA000382), anti-Ctip2 antibody (Abcam, ab18465) and anti-GFP antibodies (Nacalai tesque, Japan, 04404-26; Medical & Biological Laboratories, Japan, 598). After incubation with secondary antibodies and Hoechst 33342, the sections were washed and mounted.
Toda T., Shinmyo Y., Dinh Duong T.A., Masuda K, & Kawasaki H. (2016). An essential role of SVZ progenitors in cortical folding in gyrencephalic mammals. Scientific Reports, 6, 29578.
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2

Protein extraction and western blot analysis

Cited 1 time
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Whole-cell extracts were prepared with transgenic buffer (20 mM Tris-HCl at pH 7.5, 137 mM NaCl, 1mM EGTA, 1% Triton X-100, 10% glycerol, 1.5 mM MgCl2) supplemented with protease inhibitor cocktail (Sigma, P2714). Nuclear extracts were prepared as described previously (van den Berg et al. 2010 (link)). Western blotting was carried out according to a standard protocol (see the Supplemental Material). The antibodies used were anti-Flag (Sigma, F1804), anti-Elf5 (Santa Cruz Biotechnology, sc-9645), anti-Eomes (Abcam, ab23345; R&D Systems, MAB 6166), anti-Tfap2c (R&D System, AF5059), anti-tubulin (Abcam ab6160), and ImmunoCruz IP/WB Optima System C (sc-45040), E (sc-45042), or A (sc-45038).
Latos P.A., Sienerth A.R., Murray A., Senner C.E., Muto M., Ikawa M., Oxley D., Burge S., Cox B.J, & Hemberger M. (2015). Elf5-centered transcription factor hub controls trophoblast stem cell self-renewal and differentiation through stoichiometry-sensitive shifts in target gene networks. Genes & Development, 29(23), 2435-2448.
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3

RIP and ChIP Assays for RNA and Chromatin Analyses

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RIP assay was performed using the Magna RIP kit (Millipore) according to manufacturer’s instruction. Cell lysate from 10 million cells and 2–5ug of control IgG or antibody against SOX17 (R&D, AF1924), EOMES (Abcam, ab23345) or SMAD2/3 (R&D, AF3797) were used. RNA was extracted using phenol/chloroform and quantified by RT-qPCR. We had validated the RIP-assay using SNRNP70 antibody, which could bind to U1 snRNA.
Chromatin immunoprecipitation assay was performed using magnetic beads. Cultured cells were crosslinked using 1% formaldehyde and harvested nuclei were fragmented by sonication. Equal amount of chromatin from each sample (∼ 2 million cells each IP) and 1ug control IgG or antibody against SMAD2/3 were used. DNA was recovered by phenol/chloroform extraction. Quantitative PCR was carried out and the result was normalized to input. All primers were listed in Table S4.
Jiang W., Liu Y., Liu R., Zhang K, & Zhang Y. (2015). LncRNA DEANR1 facilitates human endoderm differentiation by activating FOXA2 expression. Cell reports, 11(1), 137-148.
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