重组Anti-NeuroD1抗体[EPR17084]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling NeuroD1 with ab205300 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic and nuclear staining on SH-SY5Y cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab205300 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Y79 (Human retinoblastoma cell line) cells labeling NeuroD1 with ab205300 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic and nuclear staining on Y79 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab205300 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NeuroD1 antibody [EPR17084] (AB205300)

IHC image of NeuroD1 staining in a section of frozen normal mouse brain performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab205300, 10 ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody. 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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh Mouse liver tissue labelling NeuroD1 with ab205300 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary at 1/1000 dilution. No staining on mouse liver tissue is observed. The nuclear counterstain was DAPI (Blue). Negative control (PMID : 24309898).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Negative tissue image : IHC image of NeuroD1 staining in a section of frozen normal mouse liver performed on a Leica Biosystems BOND® RX instrument. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab205300, 10 ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody. 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.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh Mouse hippocampus tissue labelling NeuroD1 with ab205300 at 1/50 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary at 1/1000 dilution. Positive staining on mouse hippocampus tissue is observed. The nuclear counterstain was DAPI (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling NeuroD1 with primary antibody anti-NeuroD1 (ab205300) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining in part of mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Anti-MAP2 mouse monoclonal antibody (ab11267) was used to counterstain at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) as a secondary counterstain antibody at 1/1000 dilution. The nuclear counter stain is DAPI (blue). The negative controls are as follows : - Negative control 1 : ab205300 at 1/100 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution. Negative control 2 : ab11267 Anti-MAP2 mouse monoclonal antibody at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NeuroD1 antibody [EPR17084] (ab205300) at 1/5000 dilution

All lanes:

Y79 (Human retinoblastoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/1000 dilution

Predicted band size: 40 kDa

Observed band size: 47 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-NeuroD1 antibody [EPR17084] (AB205300)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-NeuroD1 antibody [EPR17084] (ab205300) at 1/1000 dilution

Lane 1:

Human cerebellum lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Lane 4:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 40 kDa

Observed band size: 47 kDa

false

Exposure time: 3min