- PA5-29140 - Invitrogen Antibodies TDG antibody

Müller N, Ponkkonen E, Carell T, Khobta A
International journal of molecular sciences 2021 Oct 13;22(20)

Kolendowski B, Hassan H, Krstic M, Isovic M, Thillainadesan G, Chambers AF, Tuck AB, Torchia J
Epigenetics & chromatin 2018 Jan 29;11(1):5

Lühnsdorf B, Epe B, Khobta A
The Journal of biological chemistry 2014 Aug 8;289(32):22008-18

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TDG using 30 µg of A) 293T (B) A431 and C) HeLa lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a TDG polyclonal antibody (Product # PA5-29140) at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDG was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDG Polyclonal Antibody (Product # PA5-29140) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on modified whole cell extracts (1%SDS) (30 µg lysate) of HeLa (Lane 1), SW480 (Lane 2), PANC-1 (Lane 3), MCF7 (Lane 4), HCT116 (Lane 5), HT-29 (Lane 6) and HEK-293 (Lane 7). The blot was probed with Anti-TDG Polyclonal Antibody (Product # PA5-29140, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 55, 60 kDa band corresponding to TDG was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDG was performed by separating 10 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDG Polyclonal Antibody (Product # PA5-29140) at a dilution of 1:10000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using TDG Polyclonal Antibody (Product # PA5-29140). Non-transfected (–) and transfected (+) 293T whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with TDG Polyclonal Antibody (Product # PA5-29140) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using TDG Polyclonal Antibody (Product # PA5-29140). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with TDG Polyclonal Antibody (Product # PA5-29140) diluted at 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of TDG was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a TDG Polyclonal Antibody (Product # PA5-29140) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of TDG was achieved by transfecting HeLa cells with TDG specific siRNAs (Silencer® select Product # s13949). Western blot analysis (Fig. a) was performed using whole cell extracts (with 1% SDS) from TDG knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with TDG Polyclonal Antibody (Product # PA5-29140, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Reduction of signal upon siRNA mediated knock down confirms that antibody is specific to TDG.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: TDG Polyclonal Antibody detects TDG protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: TDG protein stained by TDG Polyclonal Antibody (Product # PA5-29140) diluted at 1:1,000. Blue: Hoechst 33342 staining.

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Global analysis of E2-dependent TDG localization. a MCF7 cells were treated with 100 nM of E2 (45 min) and ChIP-qPCR was performed using TDG antibody. Region used as negative control shows low level of TDG binding in ChIP-Seq data with no change in levels after E2 treatment. (* p value < 0.05, error bars represent standard deviation of the mean, n > 2). b Sites of E2-dependent TDG binding were mapped to the annotated genome using CEAS. c Sites of ER binding (+/- 1000 bp) overlaid with TDG binding signal, showing strong relationship between location of TDG binding and ER binding at these regions

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Effect of TDG knockout on the activity of GC box containing 5-mC/5-hmC/5-fC/5-caC. ( a ) Strategy for TDG knockout in Hela cells. The TDG gene was targeted simultaneously at two different sites by a pair of single guide RNAs enclosing the codon for the catalytic R140 residue. The Cas9 cut sites are indicated by arrowheads. ( b ) Validation of TDG negative clones by Western blotting. Clone F3 further used as a transfection host for the gene expression analyses is marked (*). ( c ) Expression time course of constructs containing 5-mC/5-hmC/5-fC/5-caC in the purine-rich (left group of plots) or the pyrimidine-rich GC box strand (right). Isogenic clonal TDG knockout (upper row) and NTH1 knockout (lower row) cell lines were compared with the parental HeLa cell line (overlaid in plots). All cell lines were transfected in parallel with the same sets of reporter constructs. Results of n = 3 independent experiments (mean +- SD).

PA5-29140