The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa (predicted molecular weight: 50 kDa).
Use a concentration of 1 µg/ml.
Use at an assay dependent concentration.
FunctionPossesses single-stranded DNA-stimulated ATPase and ATP-dependent DNA helicase (5' to 3') activity. Component of the NuA4 histone acetyltransferase complex which is involved in transcriptional activation of select genes principally by acetylation of nucleosomal histones H4 and H2A. This modification may both alter nucleosome - DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. This complex may be required for the activation of transcriptional programs associated with oncogene and proto-oncogene mediated growth induction, tumor suppressor mediated growth arrest and replicative senescence, apoptosis, and DNA repair. The NuA4 complex ATPase and helicase activities seem to be, at least in part, contributed by the association of RUVBL1 and RUVBL2 with EP400. NuA4 may also play a direct role in DNA repair when recruited to sites of DNA damage. RUVBL2 plays an essential role in oncogenic transformation by MYC and also modulates transcriptional activation by the LEF1/TCF1-CTNNB1 complex. May also inhibit the transcriptional activity of ATF2.
Tissue specificityUbiquitously expressed. Highly expressed in testis and thymus.
Sequence similaritiesBelongs to the ruvB family.
DomainThe C-terminal domain is required for association with ATF2.
Post-translational modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localizationNucleus matrix. Nucleus > nucleoplasm. Cytoplasm. Membrane. Mainly localized in the nucleus, associated with nuclear matrix or in the nuclear cytosol. Although it is also present in the cytoplasm and associated with the cell membranes.
ICC/IF image of ab36569 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab36569, 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 to quench autofluorescence.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).
Image courtesy of Human Protein Atlas . ab36569 staining Reptin/TIP49B/RUVB2 in human gall bladder, showing a predominantly nuclear staining pattern in glandular cells. Paraffin embedded human gall bladder tissue was incubated with ab36569 (1/400 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab36569 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues. Further results for this antibody can be found at www.proteinatlas.org
Western blot - Reptin/TIP49B/RUVB2 antibody (ab36569)
All lanes : Anti-Reptin/TIP49B/RUVB2 antibody (ab36569) at 1 µg/ml
Lane 1 : Hela cells treated with reptin-specific siRNA#1, whole cell lysate Lane 2 : Hela cells treated with reptin-specific siRNA#2, whole cell lysate Lane 3 : Hela cells treated with non-specific siRNA, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary Alexa Fluor® 680 conjugated goat anti-rabbit
at 1/7000 dilution