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Brain tissues were blocked for 60 min prior to applying goat anti-mouse serum albumin antibody conjugated to FITC (1∶300, Alpha Diagnostic International).
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Tissues were blocked for 1 h.
Tissues were blocked for 2 h with 5% normal donkey serum (NDS) in PBS containing 0.3% (retinal/brain sections) or 3% (flat mounts) Triton X-100 (PBS-TX).
Briefly, following rapid removal of the brain, the tissue was blocked coronally to contain the VTA and substantia nigra.
The brain tissues were then incubated in a blocking solution of 10% normal goat serum (NGS) and 0.3% Triton X-100 in PBS for 60 min at RT. Mouse monoclonal antibody GM3 (1∶100, NeuAc, Cosmo Bio Co., LTD, Tokyo, Japan) with 1.5% NGS, and 0.3% Triton X-100 in PBS was applied on the slides overnight at 4°C.
The brain tissues were cut equally spaced (thickness 2 mm) coronal blocks, followed by sections sliced into 10 µm in a cryostat.
Autoradiographic images of [I]AB-MECA competition under baseline and blocking conditions on rat brain tissues are shown in Fig. 4.
In our study, we found the high affinity site of etorphine (0.07 nM) in MOP/NOP KO brain tissue to be blocked by the KOP receptor antagonist, norBNI, and the low affinity site (134 nM) to be blocked by the DOP receptor antagonist, naltrindole.
For the remaining two cases, only formalin-fixed, paraffin-embedded blocks of brain tissue were available.
No blocks of brain tissue were included in the accession, and no such material exists elsewhere at the Museum.
Diagnostic tissue blocks from a non-neoplastic CP cyst and from malignant brain tissue were reviewed for tissue integrity and degree of differentiation/atypia by the paediatric neuropathologist prior to selecting blocks for sectioning and antibody staining.
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