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NDUFS3 Recombinant Rabbit Monoclonal Antibody [JU10-32]

Usd: 385

Specification

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Catalog# ET1706-07

NDUFS3 Recombinant Rabbit Monoclonal Antibody [JU10-32]

Application

  • WB

  • IHC-P

  • IF-Cell

  • IF-Tissue

  • IP

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

NDUFS3 Recombinant Rabbit Monoclonal Antibody [JU10-32]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human NDUFS3 aa 215-264 / 264.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, IF-Tissue, IP

Molecular Weight

Predicted band size: 30 kDa

Positive Control

HepG2 lysates, MCF-7, human colon carcinoma tissue, human kidney tissue.

Conjugation

unconjugated

Clone Number

JU10-32

RRID

AB_3070633

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500

  • IHC-P

  • 1:50

  • IF-Cell

  • 1:50

  • IP

  • 1:10-1:50

Target

Function

NDUFS3 (NADH dehydrogenase ubiquinone iron-sulfur protein 3) is one of about 45 subunits comprising complex I of the oxidative phosphorylation electron transport chain. The multisubunit NADH: ubiquinone oxidoreductase (complex I) is the first enzyme complex in the electron transport chain of the mitochondria. NDUFS3 is the last subunit of the seven subunits that make up the core of complex I. Through use of chaotropic agents, complex I can be separated into three different fractions: a flavoprotein fraction, an iron-sulfur protein (IP) fraction, and a hydrophobic protein (HP) fraction. The IP fraction includes NDUFS1-7. NDUFS3 contains a highly conserved casein kinase II phosphorylation site. Mutations in the NDUFS3 gene may cause optic atrophy, Leigh syndrome and complex I deficiency.

Background References

1. Kazak L et al. UCP1 deficiency causes brown fat respiratory chain depletion and sensitizes mitochondria to calcium overload-induced dysfunction. Proc Natl Acad Sci U S A 114:7981-7986 (2017).

2. Nam M et al. Mitochondrial retrograde signaling connects respiratory capacity to thermogenic gene expression. Sci Rep 7:2013 (2017).

Sequence Similarity

Belongs to the complex I 30 kDa subunit family.

Subcellular Location

Mitochondrion inner membrane.

UNIPROT

SwissProt: O75489 Human

Synonyms

CI 30 antibody

CI 30KD antibody

CI-30kD antibody

Complex I 30KD antibody

Complex I 30kDa subunit antibody

COMPLEX I, MITOCHONDRIAL RESPIRATORY CHAIN, 30-KD SUBUNIT antibody

Complex I-30kD antibody

mitochondrial antibody

NADH coenzyme Q reductase antibody

NADH dehydrogenase (ubiquinone) Fe S protein 3 30kDa antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of NDUFS3 on different lysates with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-NDUFS3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 30 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 40 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of NDUFS3 on different lysates with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-NDUFS3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 30 kDa
    Observed band size: 30 kDa

    Exposure time: 40 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1706-07) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of NDUFS3 on HepG2 lysates with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/500 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 20 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 2 minutes;<br /><br />15% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NDUFS3 on HepG2 lysates with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/500 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 20 kDa
    Observed band size: 25 kDa

    Exposure time: 2 minutes;

    15% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1706-07) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of MCF-7 cells labeling NDUFS3 with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of MCF-7 cells labeling NDUFS3 with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of MCF-7 cells labeling NDUFS3 with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of MCF-7 cells labeling NDUFS3 with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-07) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-NDUFS3 antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1706-07" style="font-weight: bold;text-decoration: underline;">ET1706-07</a>) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-NDUFS3 antibody (ET1706-07) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1706-07) at 1/50 dilution for 0.5 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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Citation

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metafield image

NDUFS3 Rabbit Polyclonal Antibody

Application: WB,IHC-P,FC

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated