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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Tet2.
Our Abpromise guarantee covers the use of ab94580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration. PubMed: 24077167|
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 224 kDa (predicted molecular weight: 224 kDa). Abcam recommends using milk (2-5%) as the blocking agent.|
Based on this data we recommend using milk as the blocking agent. We welcome customer feedback and would appreciate any comments regarding this product and the data presented above.
ICC/IF image of ab94580 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab94580 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in HeLa pfa fixed (4%, 10 minutes) cell types at 10ug/ml.
ab94580 staining Tet2 in Marmoset adult testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% milk for 30 minutes at 25°C; antigen retrieval was by heat mediation in Dako antigen retrieval solution. Samples were incubated with primary antibody (1/100 in 5% milk) for 18 hours at 4°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit polyclonal (1/500) was used as the secondary antibody.