The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 51 kDa).
SNRP70 is an RNA-binding protein that is a specific component of the U1 small nuclear ribonucleoprotein complex and constitutes the major anti-(U1) RNP autoimmune antigen. SNRP70 contains 1 RRM (RNA recognition motif) domain and mediates the splicing of pre-mRNA by binding to the loop I region of U1-snRNA.
ICC/IF image of ab83306 stained HeLa cells. The cells were 4% PFA 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 (ab83306, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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% PFA fixed (10 min) HepG2, and Hek293 cells at 5µg/ml.
IHC image of Ab83306 staining in Human Cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Ab83306, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-SNRP70 antibody (ab83306)
All lanes : Anti-SNRP70 antibody (ab83306) at 1 µg/ml
Lane 1 : Testis (Rat) Tissue Lysate Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 3 : Testis (Mouse) Tissue Lysate Lane 4 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution