Resource Report

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• ENCODE PROJECT External validation for lot: 1 is available under ENCODE ID: ENCAB501IJS - ENCODE https://www.encodeproject.org/antibodies/ENCAB501IJS

Discontinued
Discontinued at Thermo Fisher Scientific

Possibly Non-Specific
Extracted Antibody Information: "Testis extracts were prepared by mechanically disrupting testes in lysis buffer (20 mM Tris-HCL, 135 mM NaCl, 10% glycerol, 10% Nonidet P-40, 5 mM EDTA, 1 mM PMSF and 1x Complete protease inhibitor). Extracts were incubated in lysis buffer for 30 min at 4°C, spun for 10 min, and then precleared with uncoupled Protein A Dynabeads (Invitrogen, Waltham, MA) for 45 min at 4°C. Testis extracts were then added to Protein A Dynabeads chemically crosslinked to rabbit anti-YTHDC2 (A303-025A, (RRID:AB_10752592),"

Extracted Specificity Statement: "Although faint immunofluorescence signal was also detected in round and elongating spermatids in cross-sections of tubules stained with anti-YTHDC2, similar faint fluorescence was also often observed in controls stained only with secondary antibody, suggesting background. The strong signal visible in interstitial cells between testis tubules (Figure 1D) was also present in sections of testis from Ythdc2 null mutants stained with YTHDC2 antibody, indicating that the staining in interstitial cells is non-specific. Staining cross-sections of human testis tubules with anti-YTHDC2 revealed an expression pattern similar to that observed in mouse, with human YTHDC2 not detected in germ cells around the periphery of the testis tubule (Figure 1G–I, arrowheads), but present in the cytoplasm of a subset of DDX4-positive germ cells positioned further inward, including pachytene spermatocytes marked by staining of the sex vesicle by γ-H2AX (Figure 1J, arrows)."

Data was mined by Antibody Watch (https://arxiv.org/pdf/2008.01937.pdf), from PMID:29087293

Possibly Non-Specific
Extracted Antibody Information: "After lysis, lysates were centrifuged for 10 min at max speed at 4°C. The supernatant was collected and diluted with an equal volume of binding/wash buffer (25 mM Tris-HCl pH7.5, 150 mM KCl, 5 mM EDTA, 0.5% NP-40, 0.5 mM DTT, 1x Complete protease inhibitor, and 100 U/mL RNasOUT). Lysates were precleared by incubation with uncoupled Protein A Dynabeads (Invitrogen) for 1 hr at 4°C. Following preclearing, 10% of the lysate was saved for input and stored at 4°C, and the remaining lysate was added to Protein A Dynabeads chemically crosslinked to rabbit anti-YTHDC2 (A303-025A, (RRID:AB_10752592)"

Extracted Specificity Statement: "Although faint immunofluorescence signal was also detected in round and elongating spermatids in cross-sections of tubules stained with anti-YTHDC2, similar faint fluorescence was also often observed in controls stained only with secondary antibody, suggesting background. The strong signal visible in interstitial cells between testis tubules (Figure 1D) was also present in sections of testis from Ythdc2 null mutants stained with YTHDC2 antibody, indicating that the staining in interstitial cells is non-specific. Staining cross-sections of human testis tubules with anti-YTHDC2 revealed an expression pattern similar to that observed in mouse, with human YTHDC2 not detected in germ cells around the periphery of the testis tubule (Figure 1G–I, arrowheads), but present in the cytoplasm of a subset of DDX4-positive germ cells positioned further inward, including pachytene spermatocytes marked by staining of the sex vesicle by γ-H2AX (Figure 1J, arrows)."

Data was mined by Antibody Watch (https://arxiv.org/pdf/2008.01937.pdf), from PMID:29087293

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Source: Antibody Registry