Anti-Neurofilament heavy polypeptide antibody (ab8135)

Immunohistological analysis of a mouse hippocampus section stained with ab8135 at a dilution 1:2,000 in red, and co-stained with a mouse mAb to myelin basic protein (MBP) at a dilution 1:5,000 in green. The blue is DAPI staining of nuclear DNA.

Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections were stained.

ab8135 staining Neurofilament heavy polypeptide in rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections).
Tissue was fixed with formaldehyde, blocked with 5% serum for 1 hour at 25°C and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/500 in diluent) for 12 hours at 25°C. An Alexa Fluor^® 555 goat anti-rabbit polyclonal IgG (1/300) was used as the secondary antibody.

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Immunocytochemical staining of 33B cells (Rat oligodendroglioma) with ab8135 at 1/1000.

Cells were cultured in DMEM/10% FCS on coverslips in 12 well plates. Cells were washed with PBS and fixed in 4% PFA until staining. Cells were washed teice for 2 minutes and then treated with 3% hydrogen peroxide in methanol for 15 minutes at room temperature. The sample was blocked in 10% goat serum in 1% PBSA and then incubated with the primary antibody in 1% PBSA for 2 hours at room temperature. A biotinylated goat anti-rabbit was used as the secondary at 1/200 and then incubated with ABC for 30 minutes at room temperature, followed by DAB in PBS (brown). The sample was counter-stained with hematoxylin.

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ab8135 at a 1/500 dliution staining rat aged muscle tissue sections by IHC-Fr.

The tissue was permeabilized with Triton X-100 and blocked with 10% serum for 30 minutes at 25°C. Then incubated with the primary antibody for 1 hour at 25°C. Muscle was stained without fixation and post-fixed in 10% NBF following staining for 1 hour.

Green: alpha bungarotoxin stained post-synaptic neuromuscular junction.

Red: ab8135 staining Neurofilament heavy polypeptide

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