Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within RGS9. The exact sequence is proprietary.
WB: Y79 and PC12 cell lysates and human and mouse spleen tissue lysates.
ICC/IF: Neuro-2a cells.
Flow Cyt: PC12 and Y79 cells.
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 77 kDa).
1/30 - 1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/50 - 1/100.
Is unsuitable for IHC-P or IP.
Inhibits signal transduction by increasing the GTPase activity of G protein alpha subunits thereby driving them into their inactive GDP-bound form. Binds to G(t)-alpha. Involved in phototransduction; key element in the recovery phase of visual transduction.
Highly expressed in the caudate and putamen, lower levels found in the hypothalamus and nucleus accumbens and very low levels in cerebellum. Not expressed in globus pallidus or cingulate cortex. Isoform 2 is expressed predominantly in pineal gland and retina. Isoform 3 is expressed in retina (abundant in photoreceptors).
Defects in RGS9 are a cause of prolonged electroretinal response suppression (PERRS) [MIM:608415]; also known as bradyopsia. PERRS is characterized by difficulty adjusting to sudden changes in luminance levels mediated by cones.
Contains 1 DEP domain. Contains 1 G protein gamma domain. Contains 1 RGS domain.
In photoreceptor cells the DEP domain is essential for targeting RGS9 to the outer rod segments.
Retinal isoform 3 is light-dependent phosphorylated at 'Ser-478'. Phosphorylation is decreased by light exposition.
Membrane. Isoform 3 is targeted to the membrane via its interaction with RGS9BP.
Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling RGS9 with purified ab108975 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
Flow Cytometry analysis of Y79 cells labelling RGS9 with purified ab108975 at a dilution of 1/60 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Western blot - Anti-RGS9 antibody [EPR2873] (ab108975)
All lanes : Anti-RGS9 antibody [EPR2873] (ab108975) at 1/1000 dilution (unpurified)
Lane 1 : PC12 lysates Lane 2 : Human spleen lysates
Lysates/proteins at 10 µg per lane.
Secondary HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size : 77 kDa Observed band size : 55 kDa (why is the actual band size different from the predicted?) Note: Predicted molecular weights for UniProt ID O75916 isoforms 1 to 4 are ~77, 55, 57 and 50 kDa respectively. Therefore, the observed WB band may correspond to isoform 2, 3 or 4.
Vogt A et al. Isotope coded protein labeling coupled immunoprecipitation (ICPL-IP): a novel approach for quantitative protein complex analysis from native tissue. Mol Cell Proteomics12:1395-406 (2013).
Read more (PubMed: 23268931) »