PA5-40526

antibody from Invitrogen Antibodies

Targeting: NKX2-2 NKX2.2, NKX2B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-40526

Provider

Invitrogen Antibodies

Product name

Nkx2.2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Peptide sequence: MSLTNTKTGF SVKDILDLPD TNDEEGSVAE GPEEENEGPE PAKRAGPLGQ

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of human spleen cells using an anti-NKX2.2 polyclonal antibody (Product # PA5-40526).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Nkx2.2 Polyclonal Antibody (Product # PA5-40526) and a 30 kDa band corresponding to NKX2.2 was observed across cell lines and tissues tested. Due to glycosylation a 45 kDa band was also observed in cell lines which is reported. Modified whole cell extracts (1% SDS) (30 µg lysate) of Neural Stem Cell (Lane 1), U-2 OS (Lane 2), U-87 MG (Lane 3), A549 (Lane 4), tissue extracts of Mouse Pancreas (Lane 5) and Mouse Brain (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036), 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

Immunofluorescence analysis of Nkx2.2 was performed using 70% confluent log phase Neuronal stem cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Nkx2.2 Rabbit Polyclonal Antibody (Product # PA5-40526) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image showing Nucleus localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of human cells using an anti-NKX2.2 polyclonal antibody (Product # PA5-40526).