Recombinant Anti-Arc antibody [EPR18950] - BSA and Azide free (ab224717)

False colour image of Western blot: Anti-Arc antibody [EPR18950] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab183183 was shown to bind specifically to Arc. A band was observed at 45 kDa in wild-type Neuro-2a cell lysates with no signal observed at this size in arc knockout cell line ab280071 (knockout cell lysate ab280130). To generate this image, wild-type and arc knockout Neuro-2a cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Arc was immunoprecipitated from 0.35 mg of mouse brain lysate with ab183183 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab183183 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Mouse brain lysate 10 µg (Input). 

Lane 2: ab183183 IP in mouse brain lysate. 

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183183 in mouse brain lysate.

Exposure time: 1 second

Blocking/dilution buffer: 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183183).

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Arc with ab183183 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on neurons of mouse cerebral cortex. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183183).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Arc with ab183183 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Sparse nuclear staining (arrows) in the dentate gyrus of mouse hippocampus (PMID: 19750198, PMID: 19628007). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183183).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.