Alexa Fluor® 647 Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (AB317599)

ab317599 staining Anti-160 kD Neurofilament Medium in Hek-293T cells, with negative expression in NEFM knockout Hek-293T cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab317599 at 1 μg/ml (shown in green). ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488) (shown in magenta) was used at 2 μg/ml for structural counterstaining.

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This product also works with 100% methanol (5 min) fixation under the same testing conditions.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Alexa Fluor® 647 Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (AB317599)

Negative immunofluorescence staining of 160 kD Neurofilament Medium in a section of frozen mouse normal liver.

The section was fixed using 10% formaldehyde in 1X PBS for 10 minutes. No antigen retrieval step was performed prior to staining. Performed on a Leica BOND™. The section was incubated at room temperature for 1 hour with ab317599 at 1/100 dilution. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Alexa Fluor® 647 Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (AB317599)

Immunofluorescence staining of 160 kD Neurofilament Medium in a section of frozen mouse normal cerebellum.

The section was fixed using 10% formaldehyde in 1X PBS for 10 minutes. No antigen retrieval step was performed prior to staining. Performed on a Leica BOND™. The section was incubated at room temperature for 1 hour with ab317599 at 1/100 dilution (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.