Anti-NeuroD1 antibody [3H8] - BSA and Azide free

• IHC-P

AbReview50326****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (AB60704)

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 25°C. The secondary antibody used was a polyclonal goat anti-mouse/rabbit HRP conjugate, used undiluted.

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This image is courtesy of an anonymous Abreview.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (AB60704)

ab60704 at 3ug/ml staining NeuroD1 in human ovary, clear cell carcinoma by Immunohistochemistry, Formalin-fixed Paraffin-embedded tissue.

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• Flow Cyt

Unknown

Flow Cytometry - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (AB60704)

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 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 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.
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• WB

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Western blot - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (AB60704)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (ab60704)

All lanes:

IMR-32 (Human Caucasian neuroblastoma) whole cell lysate at 50 µg

Secondary

All lanes:

Goat Anti-Mouse IgG HRP at 1/2500 dilution

Predicted band size: 40 kDa

Observed band size: 150 kDa,40 kDa

true

• WB

CiteAb

Western blot - Anti-NeuroD1 antibody [3H8] - BSA and Azide free (AB60704)

Western Blotting using Anti-NeuroD1 antibody [3H8] - BSA and Azide freeAnti-NeuroD1 antibody [3H8] - BSA and Azide free, ab60704. Publication image from Minna, J. D. et al., 2019, Nat Commun, 31324758. Legend direct from paper.

Identification of IGFBP5 as a direct transcriptional target of ASCL1. a Correlation heatmap illustrating clustered matrix of top four genes with the highest similarities with ASCL1 based on the expression profiles from three independent SCLC transcriptomic datasets. b Western blot of IGFBP5 in the CM, ASCL1, and NEUROD1 in the lysates of the indicated cell lines (HBEC34-KT and NE-lung cancer cell lines). Molecular weight is indicated as Mr (k). c Immunoblotting analyses of the IGFBP5 levels in the CM and lysates of H2081 cells transduced with the indicated shRNAs. d, e RT-PCR (n = 9, 3 independent experiments x 3 replicates) (d) and immunoblotting (e) analyses of IGFBP5 expression in HEK293T cells transfected with the indicated expression constructs. f Schematic for the various IGFBP5 luciferase reporter constructs. g Luciferase activity analysis of HEK293T cells transfected with the indicated IGFBP5 luciferase reporter constructs, together with V5-tagged GFP, ASCL1, or NEUROD1 (n = 4 replicates). h Luciferase activity of HEK293T cells transfected with the indicated deletion mutants of IGFBP5 luciferase reporter constructs, together with V5-tagged GFP, ASCL1, or NEUROD1 (n = 3 replicates). Unpaired two-tailed t-test. **P < 0.01, ***P < 0.001, n.s not significant. Error bars represent mean ± standard error of mean (s.e.m.). Source data are provided as a Source Data file

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