Anti-Dnmt3a Polyclonal Antibody 2 is a Rabbit antibody targeting Dnmt3a. Anti-Dnmt3a Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Cow)

1. Blank control (Black line): U87MG (Black).
Primary Antibody (green line): Rabbit Anti-Dnmt3a antibody (TMAB-05259)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
2. Tissue/cell: human breast carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Dnmt3a Polyclonal Antibody, Unconjugated (TMAB-05259) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
3. Tissue/cell: human gastric carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-Dnmt3a Polyclonal Antibody, Unconjugated (TMAB-05259) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
4. Paraformaldehyde-fixed, paraffin embedded (rat kidney tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Dnmt3a) Polyclonal Antibody, Unconjugated (TMAB-05259) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.
5. Sample:
Lane 1: Testis (Mouse) Lysate at 40 μg
Lane 2: NIH/3T3 (Mouse) Cell Lysate at 30 μg
Lane 3: Testis (Rat) Lysate at 40 μg
Lane 4: 293T (Human) Cell Lysate at 30 μg
Lane 5: SH-SY5Y (Human) Cell Lysate at 30 μg
Lane 6: Molt-4 (Human) Cell Lysate at 30 μg
Lane 7: HepG2 (Human) Cell Lysate at 30 μg
Primary: Anti-Dnmt3a (TMAB-05259) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kD
Observed band size: 120 kD
6. Sample:
A549 (Human) Cell Lysate at 30 μg
A431 (Human) Cell Lysate at 30 μg
Hela (Human) Cell Lysate at 30 μg
Lovo (Human) Cell Lysate at 30 μg
Primary: Anti-Dnmt3a (TMAB-05259) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 130 kD
Observed band size: 120 kD

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Dnmt3a

Human

DNMT3A

DNA(cytosine-5)-methyltransferase 3A

Required for genome-wide de novo methylation and is essential for the establishment of DNA methylation patterns during development. DNA methylation is coordinated with methylation of histones. It modifies DNA in a non-processive manner and also methylates non-CpG sites. May preferentially methylate DNA linker between 2 nucleosomal cores and is inhibited by histone H1. Plays a role in paternal and maternal imprinting. Required for methylation of most imprinted loci in germ cells. Acts as a transcriptional corepressor for ZNF238. Can actively repress transcription through the recruitment of HDAC activity.