Fixed cells were labelled with primary antibodies (rabbit anti-GlyRα1, custom made; mouse anti-gephyrin mAb7a, Synaptic Systems 147011) and secondary antibodies (Cy3-conjugated goat anti rabbit IgG and Alexa Fluor 647 goat anti mouse). For PALM counting of receptor-associated mEos2-gephyrin (Fig. 2F ), mouse spinal cord neurons were fixed and endogenous GlyRs were labelled with QDs emitting at 605 nm (primary antibody: mouse anti-GlyRα1, Synaptic Systems mAb2b; secondary antibody: anti-mouse Fab’-QD605).
Mab7a
Manufactured by Synaptic Systems
Sourced in Germany
MAb7a is a monoclonal antibody that can be used for research applications. It is designed to target a specific protein or antigen, but further details about its intended use or function are not available.
Automatically generated - may contain errors
Lab products found in correlation
Mab2b, by Synaptic Systems (1 mentions)
Chicken anti gfp, by Aves Labs (1 mentions)
Mouse anti v5 antibody, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag, by Merck Group (1 mentions)
Rabbit anti vgat antibody, by Synaptic Systems (1 mentions)
Metamorph software, by Molecular Devices (1 mentions)
Lsm 700, by Zeiss (1 mentions)
Alexa 488, by Jackson ImmunoResearch (1 mentions)
4 protocols using mab7a
1
Quantitative Visualization of Glycine Receptors
2
Antibody Immunostaining Protocol
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The following antibodies were used in this study: Chicken anti GFP (1:5000, Aves Labs Inc., USA), Mouse anti-V5 antibody (1:5000, Invitrogen, Carlsbad, USA) and (1:3000, Acris, SanDiego, USA), mouse anti-FLAG (1:5000, Sigma, Saint Louis, USA), mouse anti-actin (C4 clone) antibody (1:20000, EMD Millipore Corporation, Billerica, USA), rabbit anti-VGAT antibody (1:2000, Synaptic Systems, Göttingen, Germany), mouse anti-gephyrin antibody (mAb7a, 1:1000; or 3B11, 1:10000; Synaptic Systems, Göttingen, Germany), guinea pig anti-α1 subunit (home-made; [38 (link)], as well as a STrEP-tag Purification (IBA GmbH, Göttingen, Germany).
3
Quantifying Neuronal Protein Distributions
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Immunochemical detections of GABAAR, gephyrin, synapsin, and PKC in cultured neurons were performed as previously described (Bannai et al., 2009 (link), Niwa et al., 2012 (link)) using the following antibodies: rabbit anti-GABAAR γ2 subunit antibody (Niwa et al., 2012 (link)); mouse anti-gephyrin monoclonal antibody mAb7a (0.33 μg/ml, Synaptic Systems); mouse anti-synapsin I antibody (1:3,000, Synaptic Systems); rabbit polyclonal anti-synapsin I antibody (1:400, Merck Millipore); guinea pig anti-PKCα antibody (1 μg/ml, Frontier Institute); guinea pig anti-PKCβII antibody (1 μg/ml, Frontier Institute); and guinea pig anti-PKCγ antibody (1 μg/ml, Frontier Institute). Images were acquired on an inverted microscope equipped with oil-immersion objectives (60×, numerical aperture [NA] 1.42) and a cooled charge-coupled device (CCD) camera. All images from the same culture were acquired with the same sub-saturation exposure time. Quantification of the fluorescence signal was performed using MetaMorph software (Molecular Devices) as previously described (Bannai et al., 2009 (link), Charrier et al., 2006 (link), Lévi et al., 2004 (link), Lévi et al., 2008 (link), Niwa et al., 2012 (link)).
4
Immunohistochemistry of Synaptic Proteins
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Three-week-old wild-type and α1-KO littermates were anesthetized with pentobarbital (50 mg/kg) and perfusion-fixed with 4% paraformaldehyde solution in 0.1 M sodium phosphate buffer, pH 7.4. The brain was postfixed for 6 h, cryoprotected in 30% sucrose in PBS, and cut parasagittally with a freezing microtome. Sections were pre-treated with pepsin (0.15 mg/mL in 0.2M HCl) for 10 min at 37°C 44 (link) to unmask synaptic proteins and incubated overnight in a mixture of antibodies against the α1 subunit (rabbit; custom-made), α3 subunit (guinea pig; custom-made) and gephyrin (mouse; mAb7a, Synaptic Systems, Göttingen, Germany), followed by three washes and incubation in secondary antibodies coupled to Alexa488, Cy3 and Cy5 (Jackson Immunoresearch, West Grove, PA, USA). Images were acquired by confocal laser scanning microscopy (Zeiss LSM700, Carl Zeiss MicroImaging GmbH, Germany) using a 40x objective (N.A. 1.4). Stacks of 3–10 confocal images spaced by 0.5 μm were projected for display.
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