Nicotinic Acetylcholine Receptor gamma Antibody - BSA Free

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• Nicotinic Acetylcholine Receptor gamma Antibody

  Name: Andrea Salzinger

  Application: Immunocytochemistry

  Sample Tested: Human iPSC Motor Neurons and Human NMJ containing organoid

  Species: Human

  Verified Customer | Posted 02/06/2022

  Immunofluorescent staining against gamma subunit of Acetylcholine receptors. FastMyHC: Fast myosin heavy chain (muscle fibres), Btx: Bungarotoxin (Stains alpha subunit of AChR), AChRG: gamma subunit of AChR

  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 20 m 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, 4°C. 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.

  
  Bio-Techne Response

  This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

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