PA5-50452

antibody from Invitrogen Antibodies

Targeting: NDUFAF2 B17.2L, mimitin, MMTN, NDUFA12L

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-50452

Provider

Invitrogen Antibodies

Product name

NDUFAF2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

The antibody detects endogenous levels of total NDUFAF2 protein.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

2.6 mg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of NDUFAF2 was performed by loading (from left to right): Raji cell and human liver cancer tissue, hela cell lysates (40µg) on to a 10% SDS-PAGE gel. Proteins were transferred to a membrane and the membrane was probed with a NDUFAF2 antibody (Product # PA5-50452) at a 1/250 dilution for 10 seconds, followed by a secondary antibody at 1/8000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-NDUFAF2 Polyclonal Antibody (Product # PA5-50452) and a 20 kDa band corresponding to NADH dehydrogenase [ubiquinone] 1 alpha subcomplex assembly factor 2 was observed. Whole cell extracts (30 µg lysate) of Raji (Lane 1), PC-3 (Lane 2), HEK-293 (Lane 3), SK-BR-3 (Lane 4), HeLa (Lane 5), T98G (Lane 6), MCF7 (Lane 7) and tissue extracts (30 µg lysate) of Mouse Heart (Lane 8), Rat Heart (Lane 9) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) 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 NADH dehydrogenase [ubiquinone] 1 alpha subcomplex assembly factor 2 was performed using 70% confluent log phase Hep G2 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 45 minutes at room temperature. The cells were labeled with NDUFAF2 Polyclonal Antibody (Product # PA5-50452) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly Mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).