Western blotting and immunogold EM were carried out as described19 (link). For Western blotting, samples were resolved on 4–20% or 10% Tris-glycine gels (Novex), and the primary antibodies were diluted in PBS plus 0.1% Tween 20 and 1% BSA. BR133, BR134 and BR13510 (link) at 1:4000; AT8 (Thermo; catalogue nr. MN1020) at 1:1000; MC-128 (link) at 1:10; HT7 (Thermo; catalogue nr. MN1000) at 1:2500, Anti 4R-tau (Cosmo Bio; catalogue nr. CAC-TIP-4RT-P01) and TauC425 (link) at 1:2000; and MN42324 (link) at 1:500. For immunogold EM, primary antibodies were diluted as follows: BR133, BR134, BR135, AT8, Anti 4R, TauC4 and MN423 at 1:50; and MC-1 at 1:10. Neurohistology, immunohistochemistry, and molecular genetics were carried out as described49 (link). For immunohistochemistry, the following antibody dilutions were used: RD3 (Millipore; catalogue nr. 05-803) at 1:3000; Anti-4R at 1:100; and AT8 and AT100 (Thermo; catalogue nr. MN1060) at 1:300. The brain sections were 8 μm thick, and were counterstained with haematoxylin.
Anti 4r tau
Manufactured by Cosmo Bio
Anti 4R-tau is a laboratory reagent used to detect the presence of the 4R isoform of the tau protein in biological samples. It is a specific antibody that binds to the 4R form of tau, allowing researchers to measure and analyze this protein in their experiments.
Automatically generated - may contain errors
3 protocols using anti 4r tau
1
Tau Protein Detection Protocol
2
Tau Protein Detection Protocol
3
Characterizing Tauopathy in Neurodegenerative Diseases
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Sarkosyl-insoluble fraction proteins were prepared from frozen samples of the hippocampus, hypothalamus, and midbrain periaqueductal gray matter of the patient, the brainstem of a previously reported anti-IgLON5 case (patient 7 described in Lancet Neurology)1 (link) fulfilling the postmortem diagnostic criteria of IgLON5 tauopathy, and the prefrontal cortex of a patient with Alzheimer disease (AD). Briefly, samples were homogenized and centrifuged twice at 20,000g. The supernatants were combined and incubated with 1% N-lauroylsarcosine (wt/vol) for 1 hour and centrifuged at 100,000g to get the sarkosyl-insoluble pellets that were subjected to immunoblot.
Immunoblots were run following denaturing standard procedures, proteins were transferred to a nitrocellulose membrane, and strips were incubated with anti-pTau (AT8, Thermo-Scientific, Rockford), anti-3Rtau (Merck-Millipore, Billerica, MA), and anti-4Rtau (Cosmo Bio, Tokyo, Japan) antibodies, appropriate secondary antibodies, and the results were visualized by enhanced chemiluminescence.
Immunoblots were run following denaturing standard procedures, proteins were transferred to a nitrocellulose membrane, and strips were incubated with anti-pTau (AT8, Thermo-Scientific, Rockford), anti-3Rtau (Merck-Millipore, Billerica, MA), and anti-4Rtau (Cosmo Bio, Tokyo, Japan) antibodies, appropriate secondary antibodies, and the results were visualized by enhanced chemiluminescence.
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