PA5-40526

antibody from Invitrogen Antibodies

Targeting: NKX2-2 NKX2.2, NKX2B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

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 Sequence homology: Cow: 100%; Dog: 100%; Guinea Pig: 100%; Horse: 100%; Human: 100%; Mouse: 92%; Rabbit: 92%; Rat: 100%; Sheep: 100%

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.5 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

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.