702565

antibody from Invitrogen Antibodies

Targeting: NEUROG2 Atoh4, bHLHa8, Math4A, ngn-2, NGN2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702565

Provider

Invitrogen Antibodies

Product name

Neurogenin 2 Recombinant Rabbit Monoclonal Antibody (15H30L21)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Dog, Bovine and Rabbit.

Antibody clone number

15H30L21

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Tissue extracts (30 µg lysate) of Mouse Pup Brain (Lane 1) and Nuclear enriched extracts (30 µg lysate) of U-87 MG (Lane 2), SK-N-AS (Lane 3), HeLa (Lane 4) and Hep G2 (Lane 5). The blots were probed with Anti-Neurogenin-2 (NEUROG2) Recombinant Rabbit Monoclonal Antibody (Product # 702565, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 34 kDa band corresponding to Neurogenin-2 was observed across the tissues and cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, SH-SY5Y cells treated with retinoic acid (10 uM, 5 days) were fixed and permeabilized for detection of endogenous Neurogenin-2 using Neurogenin-2 (NEUROG2) Recombinant Rabbit Monoclonal Antibody (Product # 702565, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Neurogenin-2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Neurogenin-2. Panel e) shows untreated cells with no signal and panel f) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.