- PA5-27969 - Invitrogen Antibodies UHRF1 antibody

Submitted references MiR-9-1 Suppresses Cell Proliferation and Promotes Apoptosis by Targeting UHRF1 in Lung Cancer.

Jia CY, Xiang W, Liu JB, Jiang GX, Sun F, Wu JJ, Yang XL, Xin R, Shi Y, Zhang DD, Li W, Zuberi Z, Zhang J, Lu GX, Wang HM, Wang PY, Yu F, Lv ZW, Ma YS, Fu D
Technology in cancer research & treatment 2021 Jan-Dec;20:15330338211041191

LASP-1: a nuclear hub for the UHRF1-DNMT1-G9a-Snail1 complex.

Duvall-Noelle N, Karwandyar A, Richmond A, Raman D
Oncogene 2016 Mar 3;35(9):1122-33

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of UHRF1 using 30 µg of HeLa lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a UHRF1 polyclonal antibody (Product # PA5-27969) at a dilution of 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of UHRF1 was achieved by transfecting MCF7 with UHRF1 specific siRNAs (Silencer® select Product # s70719, s70720). Western blot analysis (Fig. a) was performed using Whole cell extracts from the UHRF1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with UHRF1 Polyclonal Antibody (Product # PA5-27969, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to UHRF1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-UHRF1 Polyclonal Antibody (Product # PA5-27969) and a 90kDa band corresponding to UHRF1 was observed along with uncharacterized bands (*) across cell lines. Whole cell extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2) and MDA-MB-231 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of UHRF1 in paraformaldehyde-fixed HeLa cells using a UHRF1 polyclonal antibody (Product # PA5-27969) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded BT483 xenograft, using UHRF1 (Product # PA5-27969) antibody at 1:250 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 LASP-1 associates with epigenetic proteins UHRF1, DNMT1, G9a and Snail1 A) LASP-1 co-immunoprecipitates with UHRF1 and DNMT1. 100 mug of the nuclear extracts from MDA-Bone-Un cells stimulated with vehicle (0 min) or 500 ng /ml of CXCL12 for 15 min were incubated with control mouse monoclonal IgG (Mock) or with mouse anti-LASP-1 antibody. LASP1-bound proteins were eluted and analyzed for endogenous LASP-1 and associated proteins UHRF1 and DNMT1. Bottom panel: 15 mug of total cell lysates from MDA-Bone-Un cells that were stimulated with vehicle (0 min) or CXCL12 for 15 min. LASP-1 lysate blot was obtained after stripping and re-probing for LASP-1 after the detection of Snail1 first. The experiment was repeated twice and the representative blot was quantified and shown. NS - Non-silenced control; KD - LASP-1 knock down. The band intensities of proteins were quantified by using Image J analysis and normalized to their respective non-stimulated control (bands in -CXCL12 lane). The fold change was given above the bands to track the level of the immunoprecipitated proteins with respect to their protein levels in the lysates. B) LASP-1 co-immunoprecipitates with G9a and Snail1. 100 mug of the nuclear extracts from MDA-Bone-Un cells stimulated with vehicle (0 min and Mock) or 500 ng /ml of CXCL12 for 15 min were incubated with control mouse monoclonal IgG (Mock) or with mouse anti-LASP-1 antibody. Bound proteins were eluted and analyzed for endogenous LASP-1 and associated G9a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. The expression and clinical significance of miR-9 to 1 and UHRF1 in NSCLC. (A) IHC assay to measure UHRF1 level in 56 NSCLC tissues and 36 adjacently normal lung tissues. (B) The correlation between UHRF1 and miR-9 to 1 level in 56 NSCLC tissues. (C) Kaplan-Meier survival analysis was used to evaluate the prognostic value of UHRF1 expression for OS of 56 NSCLC tissues. (D) Kaplan-Meier survival analysis to evaluate the prognostic value of UHRF1 together with miR-9 to 1 expression in NSCLC for OS. Abbreviations: NSCLC, nonsmall cell lung cancer; miR-9 to 1, microRNA-9 to 1; OS, overall survival; UHRF1, ubiquitin-like containing PHD and RING finger domains 1.

PA5-27969

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