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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 35 kDa (predicted molecular weight: 35 kDa).
Use a concentration of 5 µg/ml.
功能Seems to bind protein kinase C acting as an intracellular receptor to anchor the activated PKC to the cytoskeleton. May be involved in up-regulation of the activity of kinases such as PKC via binding to KRT1. Together with KRT1 and ITGB1, serves as a platform for SRC activation or inactivation. May play an important role in the developing brain and neuronal differentiation.
序列相似性Belongs to the WD repeat G protein beta family. Contains 7 WD repeats.
结构域The WD repeats domain 5 mediates interaction with TRIM63.
细胞定位Cell membrane. Located on plasma membrane of neuroblastoma NMB7 cells.
ICC/IF image of ab111621 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab111621, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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 4% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa and MCF7 cells at 5µg/ml.
Western blot - Anti-RACK1 antibody (ab111621)
All lanes : Anti-RACK1 antibody (ab111621) at 1 µg/ml
Lane 1 : Liver (Human) Tissue Lysate - adult normal tissue Lane 2 : Liver (Mouse) Tissue Lysate Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 7 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 35 kDa Observed band size : 35 kDa Additional bands at : 38 kDa (possible non-specific binding),54 kDa (possible non-specific binding).
Exposure time : 30 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab111621 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Anti-RACK1 antibody (ab111621)参考文献
has not yet been referenced specifically in any publications.