1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
Plays a central role in the regulatory network for splicing, controlling the intranuclear distribution of splicing factors in interphase cells and the reorganization of nuclear speckles during mitosis. Hyperphosphorylates RS domain-containing proteins such as SFRS1, SFRS2 and ZRSR2 on serine residues during metaphase but at lower levels during interphase. Locks onto SFRS1 to form a stable complex and processively phosphorylates the RS domain. Appears to mediate HBV core protein phosphorylation which is a prerequisite for pregenomic RNA encapsidation into viral capsids.
Isoform 2 is predominantly expressed in the testis but is also present at lower levels in heart, ovary, small intestine, liver, kidney, pancreas and skeletal muscle. Isoform 1 is only seen in the testis, at lower levels than isoform 2.
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. Contains 1 protein kinase domain.
ICC/IF image of ab90527 stained HeLa 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 ab90527 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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. This antibody also gave a positive result in methanol fixed (100%, 5min) HepG2, Hek293, MCF-7 cells at 1ug/ml, and also in formaldehyde fixed (4%, 10min) HeLa, Hek293, HepG2, MCF-7 cells at 1ug/ml.
Western blot - Anti-SRPK1 antibody (ab90527)
All lanes : Anti-SRPK1 antibody (ab90527) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg Lane 5 : Whole cell lysate from NIH 3T3 cells at 50 µg
ab90527 at 1 µg/ml detecting SRPK1 following immunoprecipitation. Detection by ECL with a 3 second exposure.
Lane 1; IP with ab90527
Lane 2; IP with an antibody to a downstream epitope of SRPK1
Lane 3; IP with control IgG
All antibodies used at at 3 µg/mg lysate for immunoprecipitation. 1 mg of lysate was used for IP and 20% of IP loaded.