Anti atrx

Manufactured by Cell Signaling Technology

Anti-ATRX is a primary antibody that specifically binds to the ATRX protein. ATRX is a chromatin remodeling protein that plays a role in regulating gene expression. This antibody can be used to detect and study the ATRX protein in various research applications.

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Lab products found in correlation

Anti smarcal1, by Cell Signaling Technology (2 mentions) Anti jarid2, by Novus Biologicals (2 mentions) Anti ezh2, by Cell Signaling Technology (2 mentions) Anti α tubulin, by Merck Group (1 mentions) Nupage, by Thermo Fisher Scientific (1 mentions) Anti daxx, by Cell Signaling Technology (1 mentions) Anti atrx, by Santa Cruz Biotechnology (1 mentions) Sc 55584, by Santa Cruz Biotechnology (1 mentions) Anti gapdh, by Abcam (1 mentions) Ab7311, by Abcam (1 mentions) Anti h4, by Abcam (1 mentions) Anti ha, by Roche (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Anti eed, by Cell Signaling Technology (1 mentions) Anti suz12, by Cell Signaling Technology (1 mentions) Anti ring1b, by Abcam (1 mentions) H2ak119u1, by New England Biolabs (1 mentions) Anti h3k27me3, by Diagenode (1 mentions) Anti h3, by Abcam (1 mentions)

4 protocols using anti atrx

SDS-PAGE and Immunoblotting Protocol

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Cell were lysed
into 2% w/v SDS (50 mM Tris, pH 6.8). Lysates were separated using
discontinuous SDS-PAGE to resolve proteins of molecular weight <
200 kDa or tris-acetate gels (NuPAGE, Invitrogen) to resolve proteins
of molecular weight > 200 kDa. Antibodies used for immunoblotting
were anti-SMARCAL1 (1/100, D3P5I; Cell Signaling), anti-DAXX (1/1000,
25C12; Cell Signaling), anti-ATRX (1/1000, D1N2E; Cell Signaling),
anti-ATRX (1/100, sc-55584; Santa Cruz Biotechnology), anti-alpha-tubulin
(1/5000, DM1A; Sigma-Aldrich), and anti-GAPDH (1/5000, ab9485; Abcam).

Goncalves T., Zoumpoulidou G., Alvarez-Mendoza C., Mancusi C., Collopy L.C., Strauss S.J., Mittnacht S, & Tomita K. (2020). Selective Elimination of Osteosarcoma Cell Lines with Short Telomeres by Ataxia Telangiectasia and Rad3-Related Inhibitors. ACS Pharmacology & Translational Science, 3(6), 1253-1264.

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Dinucleosome-Protein Interaction Assay

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WT and uH2A dinucleosomes were incubated with prewashed, magnetic, streptavidin coated beads (Fisher) for 1 h at 4 °C, and then washed five times with 250 mM NaCl, 10 mM Tris-HCl pH 7.0 and 0.01% Tween 20. 10 μg dinucleosomes (WT or uH2A) were incubated with 250 ug nuclear cell extract for 1 h at 4 °C, again washed five times with 250 mM NaCl, 10 mM Tris-HCl pH 7.0 and 0.01% Tween 20, and finally the reaction stopped by adding SDS loading buffer. Samples were analysed by SDS-PAGE and western blot. Primary antibodies used were anti-HA (Roche 3F10 11867423001) at 1:1,000 dilution, anti-ATRX (a gift from R.Gibbons) at 1:10 dilution, anti-EZH2 (Cell Signalling 5246) at 1:1,000 dilution, anti-JARID2 (Novus Biologicals NB100-2214) at 1:1,000 dilution and anti-H4 (Abcam AB7311).

Cooper S., Grijzenhout A., Underwood E., Ancelin K., Zhang T., Nesterova T.B., Anil-Kirmizitas B., Bassett A., Kooistra S.M., Agger K., Helin K., Heard E, & Brockdorff N. (2016). Jarid2 binds mono-ubiquitylated H2A lysine 119 to mediate crosstalk between Polycomb complexes PRC1 and PRC2. Nature Communications, 7, 13661.

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Western Blot Analysis of SMARCAL1 and ATRX

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Cells were lysed in protein-denaturing lysis buffer and protein was quantified using the BCA Protein Assay Kit (Pierce). Equal amounts of protein were loaded on SDS-polyacrylamide gels (3–8% Tris-Acetate for blots probing for ATRX, 4–12% bis-tris for all others), transferred to membranes, blocked, and blotted with antibodies. Antibodies used included anti-SMARCAL1 (Cell Signaling Technologies), anti-ATRX (Cell Signaling Technologies), anti-β-Actin (Cell Signaling Technologies), and anti-GAPDH (Santa Cruz Biotechnology) for equal loading control. Original blots are provided in Supplementary Figures 10–11.

Diplas B.H., He X., Brosnan-Cashman J.A., Liu H., Chen L.H., Wang Z., Moure C.J., Killela P.J., Loriaux D.B., Lipp E.S., Greer P.K., Yang R., Rizzo A.J., Rodriguez F.J., Friedman A.H., Friedman H.S., Wang S., He Y., McLendon R.E., Bigner D.D., Jiao Y., Waitkus M.S., Meeker A.K, & Yan H. (2018). The genomic landscape of TERT promoter wildtype-IDH wildtype glioblastoma. Nature Communications, 9, 2087.

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CRISPR-Mediated Gene Knockout Validation

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Cells were transfected with the pX459 vector containing sgRNA target site sequences listed in Supplementary Table 1. Thirty hour after transfection, medium containing 1.5 μg ml⁻¹ puromycin was added, and after a subsequent 48 h, cells were cultured in medium without selection until colonies had grown. Successful homozygous mutation was confirmed by PCR and sequencing using primers in Supplementary Table 1 and, where possible by immunoblotting. Antibodies used were anti-JARID2 (Novus Biologicals NB100-2214) at 1:1,000 dilution, anti-SUZ12 (Cell Signalling 3737) at 1:1,000 dilution, anti-EZH2 (Cell Signalling 5246) at 1:1,000 dilution, anti-ATRX (a gift from R. Gibbons) at 1:10 dilution, anti-EED (a gift of A. Otte) at 1:500 dilution. Controls used were H2AK119u1 (NEB 8240) at 1:1,000 dilution, anti-H3K27me3 (Diagenode pAb-069-050) at 1:1,000 dilution, anti-H3 (Abcam ab1791) at 1:10,000 dilution and anti-RING1B (a gift of H. Koseki) at 1:1,000 dilution.

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