PA5-31860

antibody from Invitrogen Antibodies

Targeting: TET3 hCG_40738, MGC22014

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Chromatin Immunoprecipitation (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-31860

Provider

Invitrogen Antibodies

Product name

TET3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: Flag-human TET3 transfected 293T. Predicted reactivity: Mouse (94%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Bovine, Xenopus

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.14 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

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

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

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)

Main image

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)

Main image

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.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

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.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

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