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NDUFB9 Recombinant Rabbit Monoclonal Antibody [JE63-09]

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Specification

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Catalog# HA721015

NDUFB9 Recombinant Rabbit Monoclonal Antibody [JE63-09]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

NDUFB9 Recombinant Rabbit Monoclonal Antibody [JE63-09]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human NDUFB9 aa 30-179/179.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 22 kDa

Positive Control

HepG2 cell lysate, Jurkat cell lysate, 293T cell lysate, mouse heart tissue lysate, mouse brain tissue lysate, mouse kidney tissue lysate, rat heart tissue lysate, rat kidney tissue, human liver carcinoma tissue, human stomach tissue, mouse heart tissue, PC-3M, HeLa.

Conjugation

unconjugated

Clone Number

JE63-09

RRID

AB_3072139

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:100-1:400

  • FC

  • 1:500-1:1,000

Target

Function

NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 9 is an enzyme that in humans is encoded by the NDUFB9 gene. NADH dehydrogenase (ubiquinone) 1 beta subcomplex subunit 9 is an accessory subunit of the NADH dehydrogenase (ubiquinone) complex, located in the mitochondrial inner membrane. It is also known as Complex I and is the largest of the five complexes of the electron transport chain. The protein encoded by this gene is an accessory subunit of the multisubunit NADH:ubiquinone oxidoreductase (complex I) that is not directly involved in catalysis. Mammalian complex I is composed of 45 different subunits. It locates at the mitochondrial inner membrane. This protein complex has NADH dehydrogenase activity and oxidoreductase activity. It transfers electrons from NADH to the respiratory chain. The immediate electron acceptor for the enzyme is believed to be ubiquinone. Alternative splicing occurs at this locus and two transcript variants encoding distinct isoforms have been identified. Initially, NADH binds to Complex I and transfers two electrons to the isoalloxazine ring of the flavin mononucleotide (FMN) prosthetic arm to form FMNH2. The electrons are transferred through a series of iron-sulfur (Fe-S) clusters in the prosthetic arm and finally to coenzyme Q10 (CoQ), which is reduced to ubiquinol (CoQH2). The flow of electrons changes the redox state of the protein, resulting in a conformational change and pK shift of the ionizable side chain, which pumps four hydrogen ions out of the mitochondrial matrix.

Background References

1. Qi L. et. al. Single-Cell Transcriptomic Analysis Reveals Mitochondrial Dynamics in Oocytes of Patients With Polycystic Ovary Syndrome. Front Genet. 2020 Apr

2. Li LD. et. al. Down-Regulation of NDUFB9 Promotes Breast Cancer Cell Proliferation, Metastasis by Mediating Mitochondrial Metabolism. PLoS One. 2015 Dec

Subcellular Location

Mitochondrion inner membrane.

UNIPROT

SwissProt: Q9Y6M9 Human

SwissProt: Q9CQJ8 Mouse

Entrez Gene: 299954 Rat

Synonyms

B22 antibody

CI B22 antibody

CI-B22 antibody

complex I B22 subunit antibody

Complex I-B22 antibody

DKFZp566O173 antibody

FLJ22885 antibody

I B22 antibody

LYR motif containing protein 3 antibody

LYR motif-containing protein 3 antibody

Expand

Images

  • Western blot analysis of NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/500 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: 293T cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 22 kDa<br />Observed band size: 22 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</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 NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (HA721015) at 1/500 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: 293T cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 22 kDa
    Observed band size: 22 kDa

    Exposure time: 2 minutes;

    12% 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 (HA721015) 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.

  • Western blot analysis of NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/500 dilution.<br /><br />Lane 1: Mouse heart tissue lysate<br />Lane 2: Mouse brain tissue lysate<br />Lane 3: Mouse kidney tissue lysate<br />Lane 4: Rat heart tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 22 kDa<br />Observed band size: 22 kDa<br /><br />Exposure time: 1 minute;<br /><br />12% 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</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 NDUFB9 on different lysates with Rabbit anti-NDUFB9 antibody (HA721015) at 1/500 dilution.

    Lane 1: Mouse heart tissue lysate
    Lane 2: Mouse brain tissue lysate
    Lane 3: Mouse kidney tissue lysate
    Lane 4: Rat heart tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 22 kDa
    Observed band size: 22 kDa

    Exposure time: 1 minute;

    12% 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 (HA721015) 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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/400 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/400 dilution for 1 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 rat kidney tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/400 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 (HA721015) at 1/400 dilution for 1 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 liver carcinoma tissue with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/400 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/400 dilution for 1 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 liver carcinoma tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/400 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 (HA721015) at 1/400 dilution for 1 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 stomach tissue with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 dilution for 1 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 stomach tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 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 (HA721015) at 1/100 dilution for 1 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 mouse heart tissue with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 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/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 dilution for 1 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 mouse heart tissue with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 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 (HA721015) at 1/100 dilution for 1 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.

  • Flow cytometric analysis of PC-3M cells labeling NDUFB9.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of PC-3M cells labeling NDUFB9.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721015, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunocytochemistry analysis of HeLa cells labeling NDUFB9 with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NDUFB9 antibody (<a href="/products/HA721015" style="font-weight: bold;text-decoration: underline;">HA721015</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HeLa cells labeling NDUFB9 with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NDUFB9 antibody (HA721015) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

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Citation