MA1-26777

antibody from Invitrogen Antibodies

Targeting: NES FLJ21841

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-26777

Provider

Invitrogen Antibodies

Product name

Nestin Monoclonal Antibody (3k1)

Antibody type

Monoclonal

Antigen

Other

Description

Recommended positive controls: Human neural progenitors and A172 cells.

Antibody clone number

3k1

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Nestin Monoclonal Antibody (3k1) (Product # MA1-26777) and a 200 kDa band corresponding to Nestin was observed in NSC and the expression was increased in NSC differentiated to neurons. Whole cell extracts (30 µg lysate) of iPSC (Lane 1), NSC (Lane 2), NSC differentiated to neurons (Lane 3), Jurkat (Lane 4), MCF7 (Lane 5) and LNCaP (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of immersion fixed human fetal neural progenitor cells using Nestin Monoclonal Antibody (3k1) (Product # MA1-26777). Dilution: 10 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of immersion fixed human fetal neural progenitor cells using Nestin Monoclonal Antibody (3k1) (Product # MA1-26777). Dilution: 10 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Nestin was performed using 70% confluent log phase Neural stem cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Nestin Monoclonal Antibody (3k1) (Product # MA1-26777) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic localization. Panel e represents iPSC cells having no expression of Nestin. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.