Anti-G-protein coupled receptor 30 antibody

• WB

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Western blot - Anti-G-protein coupled receptor 30 antibody (AB39742)

GPR30 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

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Western blot - Anti-G-protein coupled receptor 30 antibody (ab39742) at 1 µg/mL

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Human brain normal tissue lysate - membrane extract (<a href='/en-us/products/unavailable/human-brain-normal-tissue-lysate-membrane-extract-ab29456'>ab29456</a>) at 10 µg

Secondary

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Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 42 kDa

Observed band size: 55 kDa

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• ICC

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Immunocytochemistry - Anti-G-protein coupled receptor 30 antibody (AB39742)

ICC/IF image of ab39742 stained U87-MG 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 ab39742 at 5μg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.

• ICC

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Immunocytochemistry - Anti-G-protein coupled receptor 30 antibody (AB39742)

ab39742 staining cultured rat primary astrocytes by ICC/IF. The cultured cells were fixed with 4% paraformaldehyde for 5 minutes and blocked with 10% donkey serum in 0.1% PBS-0.3% TritonX for 30 minutes at 24°C. The cultured cells were then stained with ab39742 at 1/500 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43μM Hoechst and can be observed in blue. In red astrocytes can be observed (monoclonal anti GFAP).

This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury

• ICC

Lab

Immunocytochemistry - Anti-G-protein coupled receptor 30 antibody (AB39742)

ab39742 staining G-protein coupled receptor 30 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab39742 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.