Anti-68kDa Neurofilament/NF-L抗体[DA2]

• Flow Cyt

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Flow Cytometry - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

Overlay histogram showing SH-SY5Y cells stained with ab7255 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab7255, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

IHC image of ab7255 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab7255, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

ab7255 staining 68kDa Neurofilament/NF-L in rat frontal cortex sections by Immunocytochemistry/Immunofluorescence. Samples were incubated with primary antibody at 1/5000 dilution (red) and chicken polyclonal to GFAP.

• WB

Lab

Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

All lanes:

Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255) at 1/5000 dilution

Lane 1:

rat brain whole tissue lysates

Lane 2:

rat spinal cord whole tissue lysates

Lane 3:

mouse brain whole tissue lysates

Lane 4:

mouse spinal cord whole tissue lysates

Predicted band size: 62 kDa

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• WB

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Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

Lane 1:

Coomassie Blue stain

Lane 2:

Heavy neurofilament antibody

Lane 3:

Anti-160 kD Neurofilament Medium antibody [3H11] (<a href='/products/unavailable/160-kd-neurofilament-medium-antibody-3h11-ab7256'>ab7256</a>)

Lane 4:

Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (ab7255)

Lane 5:

alpha Internexin antibody

Lane 6:

GFAP antibody

All lanes:

Cytoskeletal homogenates of rat spinal cord

Predicted band size: 102 kDa,62 kDa

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• WB

CiteAb

Western blot - Anti-68kDa Neurofilament/NF-L antibody [DA2] (AB7255)

68kDa Neurofilament/NF-L western blot using anti-68kDa Neurofilament/NF-L antibody [DA2] ab7255. Publication image and figure legend from Rupprecht, A., Sittner, D., et al., 2014, PLoS One, PubMed 24523901.

ab7255 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab7255 please see the product overview.

Expression of neuronal/glial markers and UCP mRNA during neuronal differentiation.A. Representative Western blots show the time-dependent expression of neuronal (TUJ-1 for young and NF for adult neurons) and astrocyte (GFAP) markers during mESCs differentiation in culture. Gels were loaded with 20 µg protein per lane. B. Real-time PCR analysis of mESCs shows the amount of UCP2, UCP4 and UCP5 mRNA relative to mRNA amounts of the housekeeping gene GAPDH at different time points during neuronal differentiation. Each data point represents the mean value and SD of 3 independent differentiation experiments.

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