Western Blot: Nicotinic Acetylcholine Receptor epsilon Antibody [NBP2-26108] - Staining of Mouse Fetal Skeletal Muscle lysate (35 ug protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence. ...read more
***Novus Innovators Program - new species or application used on a primary antibody.*** Human 3D organoids containing neuromuscular junctions (NMJs) were tested for the untested application IF staining of Acetylcholine receptor (AChR) subunit epsilon. NMJs were visualized with Bungarotoxin (Btx) which binds to the alpha subunit of the AChRs. Muscle fibres are shown with Fast Myosin Heavy Chain (FastMyHc). Antibody against AChR epsilon (NBP2-26108) did not co-localise with Btx staining on FastMyHc+ muscle fibres (Figure AChRE). Moreover, AChR epsilon Ab appears to bind to axons, as AChR epsilon staining is comparable to NfH axonal staining. Secondary only staining did not reveal any unspecific axonal binding of the secondary antibody, demonstrating that axonal like staining is owed to AChR epsilon Ab. Neuromuscular junction (NMJ) organoids were fixed in 4% PFA for 3h and subsequently washed 3x with PBS. Cryoprotection was carried out by incubation in 30% Sucrose overnight, followed by embedding in a 1:1 mixture of 30% Sucrose:OCT and cryosection into 20m slices, collected on superfrost glass slides. For IF staining, samples were incubated for 10min with 0.25% Triton X-100 to ensure membrane permeabilization, followed by blocking with 5% bovine serum albumin, 0.25% Triton X-100 in PBS for 1h, RT to avoid unspecific binding of primary antibodies. Subsequently, samples were incubated with primary antibodies in blocking solution overnight, 4C. The following day, samples were washed 2x with PBS and 1x with PBS-T (0.1% Tween 20) and incubated with appropriate fluorescent secondary antibodies for 1h, RT. Nuclei were counter stained with DAPI for 10min, followed by subsequent washing with 2x PBS and 1x PBS-T. Samples were mounted using Fluorosave. Images were taken with Zeiss LSMZ10 confocal microscope and analysed using Fiji/ImageJ.
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FAQs for Nicotinic Acetylcholine Receptor epsilon Antibody (NBP2-26108). (Showing 1 - 1 of 1 FAQ).
I am looking to buy antibodies against most of the nicotinic receptor subunits for WB and IHC application. Could you please indicate which of your antibodies have been used by other researchers and provide publications showing their use? Thanks.
A list of our antibodies directed against he nicotinic acetylcholine receptors can be found here. Any products with publications will be indicated on the search page and can be found under the publications tab on the individual product's datasheet. Furthermore, we guarantee all of our products for the applications and species list on the datasheet. This should help narrow down your search.
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