Anti-Arc antibody [EPR18950]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR18950] (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.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR18950] (AB183183)

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.

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

• IP

Supplier Data

Immunoprecipitation - Anti-Arc antibody [EPR18950] (AB183183)

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.

All lanes:

Immunoprecipitation - Anti-Arc antibody [EPR18950] (ab183183)

Predicted band size: 42 kDa,45 kDa,75 kDa

Observed band size: 42 kDa,45 kDa,75 kDa

false

• WB

Supplier Data

Western blot - Anti-Arc antibody [EPR18950] (AB183183)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lanes 1 - 4:

Western blot - Anti-Arc antibody [EPR18950] (ab183183) at 1/1000 dilution

Lane 5:

Western blot - Anti-Arc antibody [EPR18950] (ab183183) at 1/5000 dilution

Lane 1:

Mouse hippocampus lysate at 20 µg

Lane 2:

Rat hippocampus lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate at 10 µg

Lane 5:

Human hippocampus lysate at 10 µg

Secondary

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lane 5:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-Arc antibody [EPR18950] (AB183183)

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.

All lanes:

Western blot - Anti-Arc antibody [EPR18950] (ab183183) at 1/500 dilution

Lane 1:

Wild-type Neuro-2a cell lysate at 20 µg

Lane 2:

ARC knockout Neuro-2a cell lysate at 20 µg

Lane 2:

Western blot - Mouse ARC knockout Neuro-2a cell line (<a href='/en-us/products/cell-lines/mouse-arc-knockout-neuro-2a-cell-line-ab280071'>ab280071</a>)

Lane 3:

SH-SY5Y cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Lane 5:

Mouse Brain cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

• WB

Supplier Data

Western blot - Anti-Arc antibody [EPR18950] (AB183183)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Arc antibody [EPR18950] (ab183183) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat heart lysate at 10 µg

Lane 7:

Rat liver lysate at 10 µg

Lane 8:

Rat spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

true

Exposure time: 3min