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CYB5R3 Recombinant Rabbit Monoclonal Antibody [JE63-51]

Usd: 385

Specification

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Catalog# HA721008

CYB5R3 Recombinant Rabbit Monoclonal Antibody [JE63-51]

Application

  • WB

  • IHC-P

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

CYB5R3 Recombinant Rabbit Monoclonal Antibody [JE63-51]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human CYB5R3 aa 1-150/301.

Species Reactivity

Human

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 34 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, human liver tissue.

Conjugation

unconjugated

Clone Number

JE63-51

RRID

AB_3072133

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

  • IHC-P

  • 1:1,000

Target

Function

NADH-cytochrome b5 reductase 3 is an enzyme that in humans is encoded by the CYB5R3 gene. Cytochrome b5 reductase is involved in the transfer of reducing equivalents from the physiological electron donor, NADH, via an FAD domain to the small molecules of cytochrome b5. It’s also heavily involved in many oxidation and reduction reactions, such as the reduction of methemoglobin to hemoglobin.[8] Of the two forms of NADH-cytochrome b5 reductase, the membrane-bound form exists mainly on the cytoplasmic side of the endoplasmic reticulum and functions in desaturation and elongation of fatty acids, in cholesterol biosynthesis, and in drug metabolism. The erythrocyte form is located in a soluble fraction of circulating erythrocytes and is involved in methemoglobin reduction. Mutations in the CYB5R3 gene cause methemoglobinemia types I and II. This is a rare autosomal recessive disease due to a deficiency of isoform of NADH-cytochrome b5 reductase.

Background References

1. Wei L. et. al. GGPP depletion initiates metaflammation through disequilibrating CYB5R3-dependent eicosanoid metabolism. J Biol Chem. 2020 Nov

2. Gordeuk VR. et. al. The CYB5R3(c) (.350C>G) and G6PD A alleles modify severity of anemia in malaria and sickle cell disease. Am J Hematol. 2020 Nov

Subcellular Location

Endoplasmic reticulum membrane, Mitochondrion outer membrane; Cytoplasm.

UNIPROT

SwissProt: P00387 Human

Synonyms

B5R antibody

Cyb5r3 antibody

Cytochrome b5 reductase 3 antibody

Cytochrome b5 reductase antibody

DIA1 antibody

Diaphorase 1 antibody

Diaphorase-1 antibody

NADH cytochrome b5 reductase 3 antibody

NADH-cytochrome b5 reductase 3 membrane-bound form antibody

NADH-cytochrome b5 reductase 3 soluble form antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of CYB5R3 on different lysates with Rabbit anti-CYB5R3 antibody (<a href="/products/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-CYB5R3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 60 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/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</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 CYB5R3 on different lysates with Rabbit anti-CYB5R3 antibody (HA721008) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-CYB5R3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 34 kDa
    Observed band size: 34 kDa

    Exposure time: 60 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 (HA721008) 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 CYB5R3 on different lysates with Rabbit anti-CYB5R3 antibody (<a href="/products/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 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/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</a>) at 1/1,000 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 CYB5R3 on different lysates with Rabbit anti-CYB5R3 antibody (HA721008) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 34 kDa
    Observed band size: 34 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 (HA721008) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CYB5R3 antibody (<a href="/products/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</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/HA721008" style="font-weight: bold;text-decoration: underline;">HA721008</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 liver tissue with Rabbit anti-CYB5R3 antibody (HA721008) 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 (HA721008) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"