Anti-NEFH 抗体 [EPR20020] - Chicken IgY (Chimeric)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling NEFH with ab317042 at 1/10000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human kidney.
The section was incubated with 317042 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling NEFH with ab317042 at 1/10000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human prostatic hyperplasia.
The section was incubated with 317042 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NEFH with ab317042 at 1/10000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.
The section was incubated with 317042 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling NEFH with ab317042 at 1/500 (2.098 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : MCF7 (PMID : 25985363).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling NEFH with ab317042 at 1/10000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum.
The section was incubated with 317042 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Composite multiplex immunofluorescence staining of ab279297, ab317042 and ab308439 staining NeuN, NEFH and Tau (MBD region) in Mouse Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-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 ab279297 (shown in green), ab317042 (shown in magenta) and ab308439 (shown in yellow) at 1µg/ml. Cells were then incubated with ab150161 Goat F(ab')2 Anti-Rat IgG Fc (Alexa Fluor® 488) preadsorbed, ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed and ab150084 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling NEFH with ab317042 at 1/500 (2.098 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/200 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab317042 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.

-ve control 2 : ab11267 at 1/200 dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NEFH with ab317042 at 1/10000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The section was incubated with 317042 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins

• WB

Supplier Data

Western blot - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (AB317042)

Negative control : kidney

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lanes 1-2 : 10 seconds, Lanes 3-8 : 6 seconds

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NEFH antibody [EPR20020] - Neuronal Marker - Chicken IgY (Chimeric) (ab317042) at 1/1000 dilution

Lane 1:

Human Brainstem tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Mouse cerebellum tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue lysate at 20 µg

Lane 6:

Rat cerebellum tissue lysate at 20 µg

Lane 7:

Rat brain tissue lysate at 20 µg

Lane 8:

Rat kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) (<a href='/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-ab6877'>ab6877</a>) at 1/100000 dilution

Observed band size: 200-210 kDa,124 kDa

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