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NDUFB4 Recombinant Rabbit Monoclonal Antibody [JE65-30]

Usd: 385

Specification

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Catalog# HA721112

NDUFB4 Recombinant Rabbit Monoclonal Antibody [JE65-30]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

NDUFB4 Recombinant Rabbit Monoclonal Antibody [JE65-30]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human NDUFB4 aa 1-129/129.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 15 kDa

Positive Control

K-562 cell lysate, MCF7 cell lysate, Neuro-2a cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, mouse heart tissue lysates, Rat heart tissue lysate, Rat kidney tissue lysate, human kidney tissue.

Conjugation

unconjugated

Clone Number

JE65-30

RRID

AB_3072236

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

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000

Target

Function

NADH dehydrogenase (ubiquinone) 1 beta subcomplex, 4, 15kDa is a protein that in humans is encoded by the NDUFB4 gene. The NDUFB4 protein is a subunit of NADH dehydrogenase (ubiquinone), which is located in the mitochondrial inner membrane and is the largest of the five complexes of the electron transport chain. The human NDUFB4 gene codes for a subunit of Complex I of the respiratory chain, which transfers electrons from NADH to ubiquinone. However, NDUFB4 is an accessory subunit of the complex that is believed not to be 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. 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. Murray J. et. al. The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification. J. Biol. Chem. 278:13619-13622(2003)

Subcellular Location

Mitochondrion inner membrane.

UNIPROT

SwissProt: O95168 Human

SwissProt: Q9CQC7 Mouse

Entrez Gene: 288088 Rat

Synonyms

B15 antibody

CI-B15 antibody

Complex I B15 subunit antibody

Complex I-B15 antibody

NADH dehydrogenase (ubiquinone) 1 beta subcomplex 4 15kDa antibody

NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 4 antibody

NADH ubiquinone oxidoreductase B15 subunit antibody

NADH-ubiquinone oxidoreductase B15 subunit antibody

NDUB4_HUMAN antibody

NDUFB4 antibody

Images

  • Western blot analysis of NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (<a href="/products/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/2,000 dilution.<br /><br />Lane 1: K-562 cell lysate<br />Lane 2: MCF7 cell lysate<br />Lane 3: Neuro-2a cell lysate<br />Lane 4: Mouse brain tissue lysate<br />Lane 5: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 30 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/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/2,000 dilution was used in 5% NFDM/TBST 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.

    Western blot analysis of NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (HA721112) at 1/2,000 dilution.

    Lane 1: K-562 cell lysate
    Lane 2: MCF7 cell lysate
    Lane 3: Neuro-2a cell lysate
    Lane 4: Mouse brain tissue lysate
    Lane 5: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Exposure time: 30 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 (HA721112) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (<a href="/products/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-NDUFB4 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 20 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/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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 NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (HA721112) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-NDUFB4 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa

    Exposure time: 20 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 (HA721112) at 1/1,000 dilution was used in 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 NDUFB4 on mouse heart tissue lysates with Rabbit anti-NDUFB4 antibody (<a href="/products/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 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/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NDUFB4 on mouse heart tissue lysates with Rabbit anti-NDUFB4 antibody (HA721112) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 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 (HA721112) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (<a href="/products/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 dilution.<br /><br />Lane 1: Rat heart tissue lysate<br />Lane 2: Rat kidney tissue lysate<br />Lane 3: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 15 kDa<br />Observed band size: 15 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/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NDUFB4 on different lysates with Rabbit anti-NDUFB4 antibody (HA721112) at 1/1,000 dilution.

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

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 15 kDa
    Observed band size: 15 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 (HA721112) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-NDUFB4 antibody (<a href="/products/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 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/HA721112" style="font-weight: bold;text-decoration: underline;">HA721112</a>) at 1/1,000 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 kidney tissue with Rabbit anti-NDUFB4 antibody (HA721112) at 1/1,000 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 (HA721112) at 1/1,000 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.

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