- MA5-15722 - Invitrogen Antibodies SETDB1 antibody

Submitted references Complete loss of H3K9 methylation dissolves mouse heterochromatin organization.

Montavon T, Shukeir N, Erikson G, Engist B, Onishi-Seebacher M, Ryan D, Musa Y, Mittler G, Meyer AG, Genoud C, Jenuwein T
Nature communications 2021 Jul 16;12(1):4359

Foxd3 controls heterochromatin-mediated repression of repeat elements and 2-cell state transcription.

Puri D, Koschorz B, Engist B, Onishi-Seebacher M, Ryan D, Soujanya M, Montavon T
EMBO reports 2021 Dec 6;22(12):e53180

TRIM28 is an Epigenetic Barrier to Induced Pluripotent Stem Cell Reprogramming.

Miles DC, de Vries NA, Gisler S, Lieftink C, Akhtar W, Gogola E, Pawlitzky I, Hulsman D, Tanger E, Koppens M, Beijersbergen RL, van Lohuizen M
Stem cells (Dayton, Ohio) 2017 Jan;35(1):147-157

The methyltransferase SETDB1 regulates a large neuron-specific topological chromatin domain.

Jiang Y, Loh YE, Rajarajan P, Hirayama T, Liao W, Kassim BS, Javidfar B, Hartley BJ, Kleofas L, Park RB, Labonte B, Ho SM, Chandrasekaran S, Do C, Ramirez BR, Peter CJ, C W JT, Safaie BM, Morishita H, Roussos P, Nestler EJ, Schaefer A, Tycko B, Brennand KJ, Yagi T, Shen L, Akbarian S
Nature genetics 2017 Aug;49(8):1239-1250

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SETDB1 using SETDB1 monoclonal antibody (Product # MA5-15722) in MCF-7 (1), T47D (2), HEK293 (3), JURKAT (4), NIH/3T3 (5), F9 (6), RAW246.7 (7) and COS-7 (8) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of SETDB1 was achieved by transfecting MCF7 with SETDB1 specific siRNAs (Silencer® select Products # s19111, s19110). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the SETDB1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with SETDB1 Monoclonal Antibody (5H6A12) (Product # MA5-15722, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Reduction in signal upon siRNA mediated knock down confirms that antibody is specific to SETDB1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-SETDB1 Monoclonal Antibody (5H6A12) (Product # MA5-15722) and a 180 kDa band corresponding to SETDB1 was observed in all cell and tissue lysates tested. Modified whole cell lysates (1% SDS) (30 µg lysate) of SW480 (Lane 1), MCF7 (Lane 2), T-47D (Lane 3), IMR-32 (Lane 4), and tissue lysates (30 µg lysate) of Mouse Testis (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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)
Main image
Experimental details: Western blot analysis of SETDB1 using SETDB1 monoclonal antibody (Product # MA5-15722) in MCF-7 (1), T47D (2), HEK293 (3), JURKAT (4), NIH/3T3 (5), F9 (6), RAW246.7 (7) and COS-7 (8) cell lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of LOVO cells using SETDB1 monoclonal antibody (Product # MA5-15722) (Green). Red: actin filaments have been labeled with phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: EV5 Figure Role of FOXD3 in SUV39H1-mediated regulation of MSR and MERVL STRING analysis to predict putative interaction between FOXD3, SUV39H1, SUV39H2, SETDB1 and SETDB2. FOXD3 was significantly predicted to interact with SUV39H1 and SETDB1 ( P -value < 0.01). The criteria used for predicting interaction are depicted. GFP immunoprecipitation analysis using Foxd3 cKO mESCs expressing GFP-FOXD3. Immunoblot results using antibodies against SUV39H1, SETDB1 and GFP are depicted. The experiment was repeated three times using biological replicates. Volcano plot depicting significantly upregulated and downregulated repeats in Suv39h dn ES cells compared with wild-type ES cells determined by RNA-Seq analysis of nuclear RNA. MSR (GSAT_MM), MERVL-int, L1MdA and MT2_Mm are labelled. ChIP-qPCR depicting enrichment of H3K4me3 (top panel) and H3K27me3 (bottom panel) over MERVL, MSR and L1MdA in control and Foxd3 KO cells. Actb promoter is used as negative control. Data are represented as average percentage input of 3 biological replicates. Error bars represent SEM (* P < 0.05 paired t -test). Schematic depicting the location of Foxd3 siRNAs (pink) along the Foxd3 locus (upper panel). Immunofluorescence detected by FOXD3 antibody in Foxd3i cells. Nuclei are stained with DAPI. Scale bar = 10 mum (lower left). RT-qPCR analysis of Foxd3 expression in control and Foxd3i cells. Data are represented as fold change compared with control (scrambled) after normalisation to Gapdh . N = 3 biological

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure EV5 Role of FOXD3 in SUV39H1-mediated regulation of MSR and MERVL STRING analysis to predict putative interaction between FOXD3, SUV39H1, SUV39H2, SETDB1 and SETDB2. FOXD3 was significantly predicted to interact with SUV39H1 and SETDB1 ( P -value < 0.01). The criteria used for predicting interaction are depicted. GFP immunoprecipitation analysis using Foxd3 cKO mESCs expressing GFP-FOXD3. Immunoblot results using antibodies against SUV39H1, SETDB1 and GFP are depicted. The experiment was repeated three times using biological replicates. Volcano plot depicting significantly upregulated and downregulated repeats in Suv39h dn ES cells compared with wild-type ES cells determined by RNA-Seq analysis of nuclear RNA. MSR (GSAT_MM), MERVL-int, L1MdA and MT2_Mm are labelled. ChIP-qPCR depicting enrichment of H3K4me3 (top panel) and H3K27me3 (bottom panel) over MERVL, MSR and L1MdA in control and Foxd3 KO cells. Actb promoter is used as negative control. Data are represented as average percentage input of 3 biological replicates. Error bars represent SEM (* P < 0.05 paired t -test). Schematic depicting the location of Foxd3 siRNAs (pink) along the Foxd3 locus (upper panel). Immunofluorescence detected by FOXD3 antibody in Foxd3i cells. Nuclei are stained with DAPI. Scale bar = 10 mum (lower left). RT-qPCR analysis of Foxd3 expression in control and Foxd3i cells. Data are represented as fold change compared with control (scrambled) after normalisation to Gapdh . N = 3 biological

MA5-15722

Antibody data