The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 35, 39 kDa representing two alternatively spliced forms of UNG that are differentially localized (predicted molecular weight: 33, 35 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Excises uracil residues from the DNA which can arise as a result of misincorporation of dUMP residues by DNA polymerase or due to deamination of cytosine.
Isoform 1 is widely expressed with the highest expression in skeletal muscle, heart and testicles. Isoform 2 has the highest expression levels in tissues containing proliferating cells.
Involvement in disease
Defects in UNG are a cause of immunodeficiency with hyper-IgM type 5 syndrome (HIGM5) [MIM:608106]. Hyper-IgM syndrome is a condition characterized by normal or increased serum IgM concentrations associated with low or absent serum IgG, IgA, and IgE concentrations. HIGM5 is associated with profound impairment in immunoglobulin (Ig) class-switch recombination (CSR) at a DNA precleavage step.
Belongs to the uracil-DNA glycosylase family.
Isoform 1 is processed by cleavage of a transit peptide.
All lanes : Anti-UNG antibody (ab23926) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate Lane 2 : Jurkat whole cell lysate (ab7899) Lane 3 : HeLa (Human epithelial carcinoma cell line) nuclear lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Predicted band size : 33, 35 kDa Observed band size : 35,39 kDa (why is the actual band size different from the predicted?) Additional bands at : 18 kDa (possible cleavage fragment,cross reactivity),23 kDa (possible cleavage fragment,cross reactivity),40 kDa (possible cross reactivity).
ICC/IF image of ab23926stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab23926, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
ab23926 staining UNG in human heart muscle. Paraffin embedded human heart muscle tissue was incubated with ab23926 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab23926 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.
Kitamura K et al. Uracil DNA glycosylase counteracts APOBEC3G-induced hypermutation of hepatitis B viral genomes: excision repair of covalently closed circular DNA. PLoS Pathog9:e1003361 (2013).
Read more (PubMed: 23696735) »
Bulgar AD et al. Removal of uracil by uracil DNA glycosylase limits pemetrexed cytotoxicity: overriding the limit with methoxyamine to inhibit base excision repair. Cell Death Dis3:e252 (2012).
Read more (PubMed: 22237209) »