- MA5-18108 - Invitrogen Antibodies EZH2 antibody

Submitted references Quantifying the phase separation property of chromatin-associated proteins under physiological conditions using an anti-1,6-hexanediol index.

Shi M, You K, Chen T, Hou C, Liang Z, Liu M, Wang J, Wei T, Qin J, Chen Y, Zhang MQ, Li T
Genome biology 2021 Aug 17;22(1):229

Ribavirin as a potential therapeutic for atypical teratoid/rhabdoid tumors.

Casaos J, Huq S, Lott T, Felder R, Choi J, Gorelick N, Peters M, Xia Y, Maxwell R, Zhao T, Ji C, Simon T, Sesen J, Scotland SJ, Kast RE, Rubens J, Raabe E, Eberhart CG, Jackson EM, Brem H, Tyler B, Skuli N
Oncotarget 2018 Jan 30;9(8):8054-8067

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of EZH2 in T47D cell lysate (35 µg/lane) using an EZH2 monoclonal antibody (Product # MA5-18108).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of EZH2 was achieved by transfecting Hep G2 cells with EZH2 specific siRNAs (Silencer® select Product # s4918, s4917). Western blot analysis (Fig. a) was performed using whole cell extracts from the EZH2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with EZH2 Monoclonal Antibody (144CT2.1.1.5) (Product # MA5-18108, 1:2000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 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 EZH2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), MCF7 (Lane 2), HCT 116 (Lane 3), SW480 (Lane 4), Caco-2 (Lane 5), T98G (Lane 6), PC-3 (Lane 7), DU 145 (Lane 8), Jurkat (Lane9) and Hep G2 (Lane 10). The blot was probed with Anti-EZH2 Monoclonal Antibody (144CT2.1.1.5) (Product # MA5-18108, 1:2000 dilution) and detected by chemiluminescence using Goat anti Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). An 86 kDa band corresponding to EZH2 was observed across all cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of EZH2 showing staining in the nucleus of U-251 cells using an EZH2 monoclonal antibody (Product # MA5-18108) followed by detection using a fluorescent conjugated secondary antibody (green). Cytoplasmic actin was stained with a fluorescent red phalloidin (7units/mL, 1 h at 37ºC).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of EZH2 was performed using 70% confluent log phase HCT 116 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EZH2 Mouse Monoclonal Antibody (144CT2.1.1.5) (Product # MA5-18108) at 1: 50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nucleus localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of EZH2 in formalin-fixed, paraffin-embedded human testis tissue using an EZH2 monoclonal antibody (Product # MA5-18108) followed by peroxidase conjugation of the secondary antibody and DAB staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Enrichment of endogenous EZH2 protein at specific gene loci using Anti-EZH2 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-EZH2 mouse monoclonal antibody (Product # MA5-18108, 8 ul) on sheared chromatin from 2 million HCT 116 cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over the promoters of HOXA9, CCND2 (positive) and SAT2 satellite repeats and SATa satellite alpha (negative). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

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

MA5-18108

Antibody data