Immunohistochemistry analysis of DNA Methyltransferase 3 showing positive staining in the nucleus and cytoplasm of paraffin-treated Human breast carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a DNA Methyltransferase 3 polyclonal antibody (Product # PA1-884) diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues M(1) K G D S R H L N E E E G A(14) C of mouse DNMT3b|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|Western Blot (WB)||2.0 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 4 publications below|
PA1-884 detects DNA methyltransferase 3 from human and mouse tissues and cells as well as recombinant human Dnmt3. This antibody has been used to detect both recombinant and endogenous DNMT3b and recombinant DNMT3a.
PA1-884 has been successfully used in Immunohistochemistry (paraffin) and Western blot procedures. By Western blot, this antibody detects DNMT3 from P19 nuclear extracts.
The PA1-884 immunogen is a synthetic peptide corresponding to residues 1-14 of mouse DNMT3b. This sequence is 79% conserved between human and mouse. PA1-884 immunizing peptide (Cat. # PEP-117) is available for use in neutralization and control experiments.
DNA methyltransferases methylate the 5-position of cytosine in the context of CpG dinucleotides. DNA methylation is crucial for normal embryonic development, genetic imprinting, and X-chromosome inactivation. Hypermethylation of DNA has also been implicated in tumorigenesis through improper methylation of growth regulatory gene promoter regions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
G9a/GLP Complex Maintains Imprinted DNA Methylation in Embryonic Stem Cells.
PA1-884 was used in western blot to characterize the G9a/GLP complex and how it maintains imprinted DNA methylation in embryonic stem cells
|Zhang T,Termanis A,Özkan B,Bao XX,Culley J,de Lima Alves F,Rappsilber J,Ramsahoye B,Stancheva I||Cell reports (15:77)||2016|
LSH and G9a/GLP complex are required for developmentally programmed DNA methylation.
PA1-884 was used in western blot to investigate the role of LSH and G9a/GLP in DNA methylation during development
|Myant K,Termanis A,Sundaram AY,Boe T,Li C,Merusi C,Burrage J,de Las Heras JI,Stancheva I||Genome research (21:83)||2011|
De novo DNA methyltransferase DNMT3b interacts with NEDD8-modified proteins.
PA1-884 was used in western blot to study the interaction between de novo DNA methyltransferase DNMT3b and NEDD8-modified proteins
|Shamay M,Greenway M,Liao G,Ambinder RF,Hayward SD||The Journal of biological chemistry (285:36377)||2010|
LSH cooperates with DNA methyltransferases to repress transcription.
PA1-884 was used in western blot to investigate the role of LSH during DNA methylation by DNA methyltransferases in mammals
|Myant K,Stancheva I||Molecular and cellular biology (28:215)||2008|