PA5-84240

antibody from Invitrogen Antibodies

Targeting: NDUFS2 CI-49

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-84240

Provider

Invitrogen Antibodies

Product name

NDUFS2 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: KVDDAKVSPP KRAEMKTSME SLIHHFKLYT EGYQVPPGAT YTAIEAPKGE FGVYLVSDGS SRPYRCKIKA PGFAHLAGLD

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of NDUFS2 by a NDUFS2 polyclonal antibody (Product # PA5-84240). Lane 1: Marker [kDa] 250, 130, 95, 72, 55, 36, 28, 17, 10 Lane 2: Human cell line RT-4 Lane 3: Human cell line U-251MG sp Lane 4: Human plasma (IgG/HSA depleted) Lane 5: Human liver tissue.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of NDUFS2 was performed by loading 40 µg of WT (lane 1) and NDUFS2 CRISPR KO (lane 2) HAP1 cell lysates in RIPA buffer onto a 4-15% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. NDUFS2 was detected using a NDUFS2 polyclonal antibody (Product # PA5-84240) at a dilution of 1:500 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4°C. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence of NDUFS2 was performed using HAP1 wild-type and NDUFS2 KO cells that were transfected with a green or a far red fluorescent dye, respectively. Post-transfection, WT and KO cells were mixed and plated to a 1:1 ratio on coverslips as a mosaic and incubated for 24 hrs. Cells were fixed in 4% PFA (in PBS) for 15 min; cells were permeabilized with 0.1% Triton X-100 for 10 min at RT and blocked with PBS with 5% BSA, 5% goat serum, and 0.01% Triton X-100 for 30 min. Cells were stained with the NDUFS2 polyclonal antibody (Product # PA5-84240) at a 1:100 dilution overnight at 4°C. Secondary antibody incubation was performed using 0.5 µg/mL of Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 antibody (Product # A21429) together with DAPI. Imaging was performed with a 40X oil objective and analysis was performed using Image J. Cell image represents a single focal plane; WT and KO cells are outlined with a yellow (WT) or magenta (KO) dashed line. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of NDUFS2 in HeLa cells using a NDUFS2 polyclonal antibody (Product # PA5-84240). The analysis shows localization to mitochondria.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of NDUFS2 in human heart muscle and pancreas tissues using a NDUFS2 polyclonal antibody (Product # PA5-84240). Corresponding RNA-seq data are presented for the same tissues.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of NDUFS2 in human heart muscle using a NDUFS2 polyclonal antibody (Product # PA5-84240). The analysis shows high expression.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of NDUFS2 in human pancreas using a NDUFS2 polyclonal antibody (Product # PA5-84240). The analysis shows low expression as expected.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of NDUFS2 was performed on HAP1 cell lysates. Antibody-bead conjugates were prepared by adding 2 µg of NDUFS2 polyclonal antibody (Product # PA5-84240) with 30 µL of protein A-Sepharose beads and rocked overnight at 4°C. 500 µg of lysate was incubated with an antibody-bead conjugate for 2 hours at 4°C. Following centrifugation and multiple washes, 10% starting material (SM), 10% unbound fraction (UB) and immunoprecipitated fraction (IP) were processed for immunoblot using the same antibody. Ponceau stained transfer of blot is shown. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.