- PA5-31860 - Invitrogen Antibodies TET3 antibody

Submitted references Integrin α6 signaling induces STAT3-TET3-mediated hydroxymethylation of genes critical for maintenance of glioma stem cells.

Herrmann A, Lahtz C, Song J, Aftabizadeh M, Cherryholmes GA, Xin H, Adamus T, Lee H, Grunert D, Armstrong B, Chu P, Brown C, Lim M, Forman S, Yu H
Oncogene 2020 Mar;39(10):2156-2169

Identification and validation of mRNA 3'untranslated regions of DNMT3B and TET3 as novel competing endogenous RNAs of the tumor suppressor PTEN.

Roquid KAR, Alcantara KMM, Garcia RL
International journal of oncology 2020 Feb;56(2):544-558

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of TET3 using A) 30 µg 293T whole cell lysate and B) 30 µg whole cell lysate of Flag-human TET3-transfected 293T cells. Samples were loaded onto a 5% SDS-PAGE gel and probed with a TET3 polyclonal antibody (Product # PA5-31860) at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of TET3 was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a TET3 Polyclonal Antibody (Product # PA5-31860) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody., and the signal was developed with Trident ECL plus-Enhanced

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of TET3 was achieved by transfecting HEL 92.1.7 with TET3 specific siRNAs (Silencer® select Product # s47238 and s47239). Western blot analysis (Fig. a) was performed using whole cell extracts prepared in NP-40 buffer from the TET3 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with TET3 Polyclonal Antibody (Product # PA5-31860, 1:1500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 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 TET3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of TET3 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: TET3 Polyclonal Antibody (Product # PA5-31860) diluted at 1:400. Red: phalloidin, a cytoskeleton marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: TET3 Polyclonal Antibody detects TET3 protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse colon. TET3 stained by TET3 Polyclonal Antibody (Product # PA5-31860) diluted at 1:500. Antigen Retrieval: Trilogy™M (EDTA based) buffer, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: TET3 antibody immunoprecipitates TET3 protein in IP experiments. IP Sample: cell lysate/extract of TET3 gene transfected 293T cells A. Cell lysate/extract of transfected 293T cell B. Control with 2 µg of preimmune rabbit IgG C. Immunoprecipitation of TET3 by 2 µg of TET3 antibody (Product # PA5-31860) 5% SDS-PAGE The immunoprecipitated TET3 protein was detected by TET3 antibody (Product # PA5-31860) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 miR-4465 negatively regulates the expression of endogenous DNMT3B, TET3 and PTEN. (A) RT-qPCR demonstrated the effects of increasing amounts of transfected miR-4465 on the endogenous transcript levels of DNMT3B, TET3 and PTEN. (B) Western blot analyses demonstrated the effects of miR-4465 overexpression on endogenous protein levels of DNMT4B, TET3 and PTEN. (C) Relative quantification of the protein expression levels of DNMT3B, TET3 and PTEN in western blots normalized to GAPDH (n=5). (D) Dual-luciferase assays demonstrated derepression of endogenous DNMT3B, TET3 and PTEN following co-transfection with increasing amounts of gene-specific TPs. (E-G) Dual-luciferase assays demonstrated direct targeting of (E) DNMT3B-, (F) TET3- and (G) PTEN-3'UTR by miR-4465 and rescue by gene-specific target protectors. (H) RT-qPCR demonstrated the effects of transfected miR-4465 and co-transfected gene-specific target TPs on the endogenous transcript levels of DNMT3B, TET3 and PTEN. All experiments were performed in cells maintained in 10% serum. Data presented are representative of three independent trials and expressed as the mean +- SD. * P

PA5-31860

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