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HADH Recombinant Mouse Monoclonal Antibody [D10-E7-R]

Usd: 350

Specification

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Catalog# HA601203

HADH Recombinant Mouse Monoclonal Antibody [D10-E7-R]

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

HADH Recombinant Mouse Monoclonal Antibody [D10-E7-R]

Antibody Type

Recombinant Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human HADH aa 265-314 / 314.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 34 kDa

Positive Control

HepG2 cell lysate, HT-29 cell lysate, HL-60 cell lysate, A431 cell lysate, K-562 cell lysate, human liver tissue lysate, mouse liver tissue lysate, mouse heart tissue lysate, rat liver tissue lysate, rat heart tissue lysate, HeLa, human colon carcinoma tissue, human kidney tissue, human liver tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

D10-E7-R

RRID

AB_3071922

Reactivity Data

WB IF-Cell IHC-P
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Rat
Tested Tested
Tested Tested

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:200

  • IHC-P

  • 1:200-1:1:1,000

Target

Function

Hydroxyacyl-Coenzyme A dehydrogenase also known as HADH is an enzyme which in humans is encoded by the HADH gene. This gene is a member of the 3-hydroxyacyl-CoA dehydrogenase gene family. The encoded protein functions in the mitochondrial matrix to catalyze the oxidation of straight-chain 3-hydroxyacyl-CoAs as part of the beta-oxidation pathway. Its enzymatic activity is highest with medium-chain-length fatty acids. Mutations in this gene cause one form of familial hyperinsulinemic hypoglycemia. A deficiency is associated with 3-hydroxyacyl-coenzyme A dehydrogenase deficiency.

Background References

1. "Sequestration of the active site by interdomain shifting. Crystallographic and spectroscopic evidence for distinct conformations of L-3-hydroxyacyl-CoA dehydrogenase." Barycki J.J., O\'Brien L.K., Strauss A.W., Banaszak L.J. J. Biol. Chem. 275:27186-27196(2000)

2. "Fulminant hepatic failure associated with mutations in the medium and short chain L-3-hydroxyacyl-CoA dehydrogenase gene." O\'Brien L.K., Rinaldo P., Sims H.F., Alonso E.M., Charrow J., Jones P.M., Bennett M.J., Barycki J.J., Banaszak L.J., Strauss A.W. J. Inherit. Metab. Dis. 23 Suppl. 1:127-127(2000)

3. "3-hydroxyacyl-CoA dehydrogenase and short chain 3-hydroxyacyl-CoA dehydrogenase in human health and disease." Yang S.-Y., He X.-Y., Schulz H. FEBS J. 272:4874-4883(2005)

Subcellular Location

Mitochondrion matrix.

UNIPROT

SwissProt: Q16836 Human

SwissProt: Q61425 Mouse

SwissProt: Q9WVK7 Rat

Synonyms

3 hydroxyacyl Coenzyme A dehydrogenase antibody

HAD antibody

HADH antibody

HADH1 antibody

HADHSC antibody

HADHSC, formerly antibody

HADSC, formerly antibody

HCDH antibody

HCDH_HUMAN antibody

HHF4 antibody

Expand

Images

  • Western blot analysis of HADH on different lysates with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: HT-29 cell lysate (20 µg/Lane)<br />Lane 3: HL-60 cell lysate (20 µg/Lane)<br />Lane 4: A431 cell lysate (20 µg/Lane)<br />Lane 5: K-562 cell lysate (20 µg/Lane)<br />Lane 6: Human liver tissue lysate (40 µg/Lane)<br />Lane 7: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 8: Mouse heart tissue lysate (40 µg/Lane)<br />Lane 9: Rat liver tissue lysate (40 µg/Lane)<br />Lane 10: Rat heart tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 34 kDa<br />Observed band size: 34 kDa<br /><br />Exposure time: 24 seconds;<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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of HADH on different lysates with Mouse anti-HADH antibody (HA601203) at 1/1,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: HT-29 cell lysate (20 µg/Lane)
    Lane 3: HL-60 cell lysate (20 µg/Lane)
    Lane 4: A431 cell lysate (20 µg/Lane)
    Lane 5: K-562 cell lysate (20 µg/Lane)
    Lane 6: Human liver tissue lysate (40 µg/Lane)
    Lane 7: Mouse liver tissue lysate (40 µg/Lane)
    Lane 8: Mouse heart tissue lysate (40 µg/Lane)
    Lane 9: Rat liver tissue lysate (40 µg/Lane)
    Lane 10: Rat heart tissue lysate (40 µg/Lane)

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

    Exposure time: 24 seconds;

    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 (HA601203) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) 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 HADH on different lysates with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-HADH 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: 15 seconds; ECL: K1801;<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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of HADH on different lysates with Mouse anti-HADH antibody (HA601203) at 1/2,000 dilution.

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

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 15 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 (HA601203) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling HADH with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 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 Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling HADH with Mouse anti-HADH antibody (HA601203) at 1/200 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 Mouse anti-HADH antibody (HA601203) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 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 colon carcinoma tissue with Mouse anti-HADH antibody (HA601203) at 1/200 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 (HA601203) at 1/200 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 Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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 Mouse anti-HADH antibody (HA601203) 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 (HA601203) 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 Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</a>) at 1/200 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 Mouse anti-HADH antibody (HA601203) at 1/200 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 (HA601203) at 1/200 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 liver tissue with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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 mouse liver tissue with Mouse anti-HADH antibody (HA601203) 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 (HA601203) 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 rat liver tissue with Mouse anti-HADH antibody (<a href="/products/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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/HA601203" style="font-weight: bold;text-decoration: underline;">HA601203</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 rat liver tissue with Mouse anti-HADH antibody (HA601203) 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 (HA601203) 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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