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Af1759

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AF1759 is a laboratory reagent product manufactured by R&D Systems. It is a buffer solution used in various experimental procedures.

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Lab products found in correlation

Phenol chloroform extraction, by Merck Group (1 mentions) Protein g dynabead, by Thermo Fisher Scientific (1 mentions) Goat anti mouse, by Thermo Fisher Scientific (1 mentions) Mab2018, by R&D Systems (1 mentions) Abcg2, by BD (1 mentions) Dapi mounting medium, by Thermo Fisher Scientific (1 mentions) Donkey anti goat igg, by Thermo Fisher Scientific (1 mentions) Leica tcs spe confocal microscope, by Leica Microsystems (1 mentions)

3 protocols using af1759

1

ChIP-seq Analysis of Histone Modifications

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Nuclei from 4 × 107 cells were collected as described (Ramirez-Carrozzi et al. 2006 (link)). Nuclear pellets were sonicated with a Misonix 3000 sonicator and subsequently incubated overnight with ChIP-grade antibodies for Oct4 (R&D AF1759), Sox2 (R&D AF2018), H3K27ac (Active Motif 39133), H3K27me3 (Active Motif 39155), or H3K9me3 (Abcam ab8898). Antibody-bound complexes were collected using protein G Dynabeads (Invitrogen 10004D) and then reverse-cross-linked with proteinase K (Thermo Fisher Scientific EO0491) overnight at 60°C. Immunoprecipitated DNA was purified using phenol-chloroform extraction (Sigma P3803) and quantified by Qubit (Thermo Fisher Q32854). The enrichment of chromatin fragments was measured by qPCR with primer pairs designed to generate 100- to 125-bp amplified products within ±200 bp of the genomic sites of interest. Quantification of fold enrichment was calculated based on the fold change of the percentage of input between target genomic loci and a negative control region (Hbb-b2).
Lo J.H., Edwards M., Langerman J., Sridharan R., Plath K, & Smale S.T. (2022). Oct4:Sox2 binding is essential for establishing but not maintaining active and silent states of dynamically regulated genes in pluripotent cells. Genes & Development, 36(19-20), 1079-1095.
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2

Multimarker Immunostaining Protocol

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Antibodies against human PDHA1(cell signaling, C54G1), tubulin(sigma,103M4773), CD44(BD pharmingen 559942), ABCG2(BD pharmingen,561451), Nanog(cell signaling 1E6C4), Sox2(RD, MAB2018) and Oct3/4(RD, AF1759 in addition to the APC isotype control( BD pharmingen 555745) and the second antibody goat anti-mouse(Life technology, 1484573) were applied in this study.
Zhong Y., Li X., Ji Y., Li X., Li Y., Yu D., Yuan Y., Liu J., Li H., Zhang M., Ji Z., Fan D., Wen J., Goscinski M.A., Yuan L., Hao B., Nesland J.M, & Suo Z. (2017). Pyruvate dehydrogenase expression is negatively associated with cell stemness and worse clinical outcome in prostate cancers. Oncotarget, 8(8), 13344-13356.
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3

Pluripotency Marker Immunostaining Protocol

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Cells were fixed in 4% paraformaldehyde for 20 min. Immunostaining was performed according to standard protocols. Antibodies were diluted in 1X PBS 0.1% BSA. Primary antibodies and concentration: goat anti-SOX2 1:100 (#LSBio (LifeSpan) Cat#LS-C132162-200, RRID: AB_10833714, R&D), goat anti-OCT4 1:200 (R and D Systems Cat# AF1759, RRID: AB_354975), goat anti-NANOG 1:100 (R and D Systems Cat# AF1997, RRID: AB_355097), and mouse anti-HLA-ABC 1:100 (Thermo Fisher Scientific Cat# MA5-11723, RRID: AB_10985125). Secondary antibodies and concentration: donkey anti-goat IgG (Thermo Fisher Scientific Cat# A32814TR, RRID: AB_2866497) and donkey anti-mouse IgG Alexa Fluor 555 1:500 (Thermo Fisher Scientific Cat# A-31570, RRID: AB_2536180). Nuclei were labeled with DAPI mounting medium (P36962, Thermo Fisher Scientific, Waltham, MA, USA). Stained cells were analyzed on a Leica TCS SPE confocal microscope (Leica Microsystems, Wetzlar, Germany).
Sesé B., Íñiguez-Muñoz S., Ensenyat-Mendez M., Llinàs-Arias P., Ramis G., Orozco J.I., Fernández de Mattos S., Villalonga P, & Marzese D.M. (2022). Glioblastoma Embryonic-like Stem Cells Exhibit Immune-Evasive Phenotype. Cancers, 14(9), 2070.
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