The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 0.25 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 52 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pineal tissue labelling SNRP70 with ab51266 at 1/100. A Cy3 conjugated donkey anti-rabbit IgG (1/200) was used as teh secondary antibody. Positive staining shown in the nuclei of pinealocytes. Magnification: 20X. Exposure time: 0.5 - 2.0 seconds. Left - DAPI. Middle - SNRP70. Right - Merge.
Western blot - Anti-SNRP70 antibody (ab51266)
Anti-SNRP70 antibody (ab51266) at 0.25 µg/ml (in 5% skim milk / PBS buffer) + HepG2 cell lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution
ab51266 (2µg/ml) staining SNRP70 in human liver using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear stain. Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
ICC/IF image of ab51266 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 (ab51266, 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.
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