The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/50 - 1/100.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
1/500 - 1/2000. Detects a band of approximately 60 kDa (predicted molecular weight: 54 kDa).
Transcriptional activator that binds to the enhancer of the adenovirus E1A gene; the core-binding sequence is 5'[AC]GGA[AT]GT-3'.
Belongs to the ETS family. Contains 1 ETS DNA-binding domain.
Sumoylated; enhanced upon ERK/MAP kinase pathway activation, it positively regulates the transcriptional activator capacity. Sumoylation at Lys-96 probably requires phosphorylation at Ser-101. Transiently polysumoylated and desumoylated by SENP1. Sumoylation is a prerequisite to polyubiquitination which in turn increases proteasomal-mediated degradation. Probably polyubiquitinated by RNF4 and deubiquitinated by USP2.
Western blot - Anti-Pea3 antibody [1A2G3] (ab70425)
Anti-Pea3 antibody [1A2G3] (ab70425) at 1/2000 dilution + Cell lysates prepared from K562 cells at 100 µg
Secondary HRP-conjugated Goat polyclonal to mouse IgG1
Predicted band size: 54 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pea3 antibody [1A2G3] (ab70425)This image is courtesy of an Abreview by Imran Ahmad.
ab70425 staining Pea3 in Human bladder transitional cell carcinoma tissue by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were fixed with formalin overnight and subjected to heat mediated antigen retrieval in citrate buffer prior to blocking with 5% BSA for 30 minutes. The primary antibody was diluted 1/200 (5% BSA in PSA) and incubated with the sample for 1 hour at 20°C. An HRP-conjugated Goat anti-Mouse polyclonal was used undiluted as the secondary antibody.
Overlay histogram showing K562 cells stained with ab70425 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab70425, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Gysin S et al. Analysis of mRNA profiles after MEK1/2 inhibition in human pancreatic cancer cell lines reveals pathways involved in drug sensitivity. Mol Cancer Res10:1607-19 (2012).
Read more (PubMed: 22833572) »
Ahmad I et al. K-Ras and ß-catenin mutations cooperate with Fgfr3 mutations in mice to promote tumorigenesis in the skin and lung, but not in the bladder. Dis Model Mech4:548-55 (2011).
Read more (PubMed: 21504907) »