The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 3 µg/ml.
Use at an assay dependent concentration. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
Use at an assay dependent concentration. The detection limit for ab60704 is approximately 0.03ng/ml as a capture antibody.
Use at an assay dependent concentration.
Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Differentiation factor required for dendrite morphogenesis and maintenance in the cerebellar cortex. Transcriptional activator. Binds to the insulin gene E-box.
Involvement in disease
Defects in NEUROD1 are the cause of maturity-onset diabetes of the young type 6 (MODY6) [MIM:606394]. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
Contains 1 basic helix-loop-helix (bHLH) domain.
Phosphorylated. In islet cells, phosphorylated on Ser-274 upon glucose stimulation; which may be required for nuclear localization. In activated neurons, phosphorylated on Ser-335; which promotes dendritic growth.
Secondary Goat Anti-Mouse IgG HRP at 1/2500 dilution
Developed using the ECL technique.
Predicted band size: 40 kDa Observed band size: 40 kDa Additional bands at: 150 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuroD1 antibody (ab60704)This image is courtesy of an anonymous Abreview.
Immunohistochemical analysis of formaldehyde fixed human cerebellum sections incubated with ab60704 for 20 minutes at 25°C in a concentration of 1/400. The blocking step was performed with 3% H2O2 for 10 minutes at 25ºC. The secondary antibody used was a polyclonal goat anti-mouse/rabbit HRP conjugate, used undiluted.
ab60704 at 3ug/ml staining NeuroD1 in human ovary, clear cell carcinoma by Immunohistochemistry, Formalin-fixed Paraffin-embedded tissue.
Immunocytochemistry/ Immunofluorescence - Anti-NeuroD1 antibody (ab60704)Image taken from an anonymous abreview
Immunofluorescence image of NeuroD1 (ab60704) on D3 mouse embryonic stem cells after 14 days of differentiation. The cells were fixed in paraformaldehyde and permeabilized in block solution. In this image you can see NeuroD1 positive staining with ab60704 in green.
Immunohistochemistry (Frozen sections) - Anti-NeuroD1 antibody (ab60704)This image is courtesy of an Abreview submitted by Ellie Tuck
ab60704 staining NeuroD1 in Mouse brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.5% Triton in TBS for 30 minutes and blocked with 5% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500) for 15 hours at 4°C. An undiluted Alexa Fluor®633-conjugated Goat anti-mouse monoclonal was used as the secondary antibody.
Overlay histogram showing SHSY-5Y cells stained with ab60704 (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 (ab60704, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.