The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 35, 81 kDa (predicted molecular weight: 81 kDa).
Use at 2-5 µg/mg of lysate.
1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
May be involved in the regulation of gene expression by covalent modification of histone proteins. Isoform Long is a corepressor of estrogen receptor (ER). Isoform Short binds to ER and sequesters it in the cytoplasm and enhances non-genomic responses of ER.
Widely expressed. High expression in brain, ovaries, adrenal glands and virgin mammary glands. Higher in tumors than in adjacent normal tissue from the same individual.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human non-small cell lung cancer tissue labelling MTA1 with ab71153 at 1/1000 (0.2 µg/ml). Detection: DAB.
Western blot - MTA1 antibody (ab71153)
All lanes : Anti-MTA1 antibody (ab71153) at 0.04 µg/ml
Lane 1 : Whole cell lysate from Hela cells at 50 µg Lane 2 : Whole cell lysate from Hela cells at 15 µg Lane 3 : Whole cell lysate from Hela cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg Lane 5 : Whole cell lysate from NIH3T3 cells at 50 µg
Predicted band size : 81 kDa Observed band size : 81 kDa Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.
Immunoprecipitation - MTA1 antibody (ab71153)
Immunoprecipitation/ Western Blot of MTA1
Lane 1: ab71153 at 3µg/mg whole cell lysate.
Lane 2: Control IgG.
Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
Subsequent WB detection was performed using 1 µg/ml ab71153.
Chemiluminescence with an exposure time of 1 second.
ICC/IF image of ab71153 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab71153, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Akiyama T et al. Transient bursts of Zscan4 expression are accompanied by the rapid derepression of heterochromatin in mouse embryonic stem cells. DNA Res22:307-18 (2015).
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