The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 64 kDa (predicted molecular weight: 58 kDa).
Use a concentration of 5 µg/ml.
Transcription factor that plays an important role in the regulation of lymphocyte differentiation. Plays an essential role in regulation of B-cell differentiation, proliferation and maturation to an effector state. Involved in regulating BCL2 expression and controlling apoptosis in T-cells in an IL2-dependent manner.
Expressed most strongly in peripheral blood leukocytes, the spleen, and the thymus.
Belongs to the Ikaros C2H2-type zinc-finger protein family. Contains 6 C2H2-type zinc fingers.
Phosphorylation on tyrosine residues induced by IL2 is required for dissociation from HRAS and nuclear translocation of IKZF3 in T-cells. Phosphorylation on tyrosine residues induced by IL4 is required for dissociation from Bcl-X(L) in T-cells.
IKZF3 was immunoprecipitated using 0.5mg Raji whole cell extract, 5µg of Rabbit polyclonal to IKZF3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Raji whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab64400.
Exposure time : 10 minutesZinc finger protein Aiolos (IKZF3) contains a number of potential phosphorylation sites (SwissProt data) which may explain it running at a higher molecular weight than predicted. IKZF3 has 6 isoforms of differing molecular weight and the 64 kDa band is thought to represent isoform 1 (58 kDa) in a phosphorylated form. The band of a slightly lower intensity seen at 48 kDa, is thought to represent isoform 2 (51 kDA).
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