Ab21990

Manufactured by Merck Group

Ab21990 is a laboratory equipment product manufactured by the Merck Group. It is designed for general laboratory use. The core function of Ab21990 is to facilitate various experimental procedures and analyses in a research or testing environment.

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Lab products found in correlation

Ab9110, by Abcam (1 mentions) Lds sample buffer, by GenScript (1 mentions) Bradford assay reagent, by Thermo Fisher Scientific (1 mentions) Immobilon p transfer membrane, by Merck Group (1 mentions) Imagequant las 4000, by GE Healthcare (1 mentions) Ripa buffer, by Boston BioProducts (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Roche (1 mentions) Norgen rna extraction kit, by Norgen Biotek (1 mentions) β actin, by Merck Group (1 mentions)

2 protocols using ab21990

Western Blot Analysis of Stem Cell Markers

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Cells were lysed in RIPA buffer (Boston BioProducts, Inc.) supplied with PMSF (Roche, 11359061001) and protease inhibitor cocktail (Sigma, P8340). Protein concentration was measured by Bradford assay reagent (Thermo Scientific). The protein amounts were adjusted among samples, then 4 × LDS sample buffer (GenScript, M00676–10) was added. Samples were boiled at 95 °C for 5 min. Proteins were separated on GenScript SurePAGETM Bis-Tris gels and blotted on the Immobilon-P transfer membrane (Millipore). The membrane was blocked with 5% skim milk and incubated with primary antibodies: anti-Nanog (1:500, rabbit IgG, Bethyl/Fisher, A300–397A), anti-Gapdh (1:1000, rabbit IgG, ProteinTech, 10494–1-AP), anti-HA tag (1:1000, rabbit IgG, Abcam, ab9110), anti-β-actin (1:1000, mouse IgG, Sigma-Aldrich, clone AC-15, A5441). anti-Otx2 (1:1000, rabbit IgG, Abcam, ab21990), anti-Prdm1 (1:1,000, mouse IgG, Sigma-Aldrich, clone 5E7, SAB5300402), anti-Tex10 (1:1000, rabbit IgG, Thermo Fisher, 720257), anti-Vinculin (1:10000, rabbit IgG, Abcam, ab129002), anti-Psmd7 (1:1000, mouse IgG, Santa Cruz, sc-390705). Then it was incubated with a secondary antibody and detected using a Medical Film Processor (SRX-101A) or ImageQuant LAS 4000 (GE Healthcare).

Li D., Yang J., Ma F., Malik V., Zang R., Shi X., Huang X., Zhou H, & Wang J. (2023). The pluripotency factor Tex10 finetunes Wnt signaling for PGC and male germline development. bioRxiv.

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Knockdown of OTX2 in Medulloblastoma Cell Lines

Cited 2 times

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OTX2 knock down in HDMB03 and MB3W1 G3 MB cells (2×10⁵ cells/well) was performed as previously described^{33 (link),34 (link)}. Briefly, OTX2 was silenced using 30 nM Silencer Select siRNAs 9931 or 9932 (Life Technologies). A non-silencing (scramble) served as the negative control. For bulk RNA sequencing, OTX2 was knocked down in three independent biological replicates for each cell line and silencing was confirmed by western blot (OTX2, Abcam, ab21990, rabbit, at 1:500, and β-actin, Sigma-Aldrich, A2228, mouse, at 1/1000 was used as a loading control) 72 hours following transfection. Total RNA was extracted from all samples using the Norgen RNA extraction kit (Norgen Biotek), and bulk RNA sequencing (RNAseq) was performed by StemCore laboratories at the Ottawa Hospital Research Institute (Ottawa, ON, Canada). For single-nucleus RNA sequencing (snRNAseq), the above was repeated but using 4.5X10⁵cells/well for MB3W1 and only one replicate was performed. Granule neuron differentiation was validated by western blot for RBFOX3 (NeuN) (Cell Signaling Technology, D4G4O, at 1:1000).

Johjima T., Itoh N., Kabuto M., Tokimura F., Nakagawa T., Wariishi H, & Tanaka H. (1999). Direct interaction of lignin and lignin peroxidase from Phanerochaete chrysosporium. Proceedings of the National Academy of Sciences of the United States of America, 96(5), 1989-1994.

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