Immunofluorescent analysis of recombinant Red Fluorescent Protein (RFP)-transfected HeLa cells. Cells were transfected with a pCMV RFP C-Myc plasmid (Product # 82026), and 48 hours post-transfection cells were fixed with formalin and permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were blocked with 3% BSA for 15 minutes at room temperature, and then probed with an anti-RFP polyclonal antibody (Product # PA1-986) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35553) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Tag|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues V(22) N G H E F E I E G E G E G R(36) of the RFP tag.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-986 detects RFP tagged proteins. This antibody is specific for the RFP tag and does not detect GFP or GFP tagged proteins.
PA1-986 has been successfully used in Western blot procedures. By Western blot, this antibody detects the presence of RFP tagged proteins. PA1-986 non-specifically detects a 45 kDa protein from E. coli extracts. This non-specific cross-reactivity is not present in mammalian samples.
PA1-986 immunogen is a synthetic peptide corresponding to residues V(22) N G H E F E I E G E G E G R(36) of the RFP tag. This peptide (Cat. # PEP-260) is available for use in neutralization and control experiments.
Epitope tagging is a powerful and versatile strategy for detecting and purifying proteins expressed by cloned genes. To utilize this feature, protein expression vectors are typically engineered with a nucleotide sequence that encodes the peptide epitope tag. The gene of interest is cloned in-frame relative to the tag and, upon expression, the protein of interest is synthesized as a fusion protein with the peptide tag. Fusion protein detection and/or purification is mediated by highly specific antibodies to the engineered peptide, thus eliminating the need for antibodies to proteins from each newly cloned gene. Commonly used epitope tags include glutathione-S-transferase (GST), c-myc, 6-histidine (6X-His), FLAG®, green fluorescent protein (GFP), red fluorescent protein (RFP, DSRed), maltose binding protein (MBP), influenza A virus haemagglutinin (HA), b-galactosidase, and GAL4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Alcohol facilitates HCV RNA replication via up-regulation of miR-122 expression and inhibition of cyclin G1 in human hepatoma cells.
PA1-986 was used in western blot to study the role of miR-122 and cyclin G1 in the mechanism by which ethanol promotes the replication of HCV RNA
|Hou W,Bukong TN,Kodys K,Szabo G||Alcoholism, clinical and experimental research (37:599)||2013|