The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Flow Cyt: 1/80.
ICC/IF: 1/100 - 1/250.
IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use of HRP conjugated or polymerized HRP secondary antibodies recommended. Stronger signals have been found using the polymerized HRP secondary.
WB: 1/25,000 - 1/50,000. Predicted molecular weight: 46 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Regulatory subunit of the cAMP-dependent protein kinases involved in cAMP signaling in cells. Type II regulatory chains mediate membrane association by binding to anchoring proteins, including the MAP2 kinase.
Four types of regulatory chains are found: I-alpha, I-beta, II-alpha, and II-beta. Their expression varies among tissues and is in some cases constitutive and in others inducible.
Belongs to the cAMP-dependent kinase regulatory chain family. Contains 2 cyclic nucleotide-binding domains.
Phosphorylated by the activated catalytic chain.
Cytoplasm. Cell membrane. Co-localizes with PJA2 in the cytoplasm and at the cell membrane.
Overlay histogram showing SH-SY5Y cells stained with ab75993 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab75993, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.