HADH Mouse Monoclonal Antibody [D10-E7]
Catalog# M1409-2
HADH Mouse Monoclonal Antibody [D10-E7]
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WB
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IF-Cell
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IHC-P
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Human
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Mouse
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Rat
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Zebrafish
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unconjugated
Overview
Product Name
HADH Mouse Monoclonal Antibody [D10-E7]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Synthetic peptide within Human HADH aa 265-314 / 314.
Species Reactivity
Human, Mouse, Rat, Zebrafish
Validated Applications
WB, IF-Cell, IHC-P
Molecular Weight
Predicted band size: 34 kDa
Positive Control
HepG2 cell lysate, HeLa 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, zebrafish tissue lysate, HeLa, human kidney tissue, human colon carcinoma tissue, human liver tissue, mouse liver tissue, rat liver tissue.
Conjugation
unconjugated
Clone Number
D10-E7
RRID
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*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:100
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IHC-P
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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. Zhu H et al. Hadh deficiency induced oligoasthenoteratozoospermia through the TNF-alpha/Bcl-2 pathway in male mice. FASEB J. 2022 Dec
2. Chu C et al. HADH suppresses clear cell renal cell carcinoma progression through reduced NRF2-dependent glutathione synthesis. Transl Oncol. 2024 Nov
Sequence Similarity
Belongs to the 3-hydroxyacyl-CoA dehydrogenase family.
Tissue Specificity
Expressed in liver, kidney, pancreas, heart and skeletal muscle.
Post-translational Modification
Succinylation at Lys-81, adjacent to a coenzyme A binding site. Desuccinylated by SIRT5 (By similarity).
Subcellular Location
Mitochondrion matrix.
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
ExpandImages
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Western blot analysis of HADH on different lysates with Mouse anti-HADH antibody (M1409-2) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: HT-29 cell lysate (20 µg/Lane)
Lane 4: HL-60 cell lysate (20 µg/Lane)
Lane 5: A431 cell lysate (20 µg/Lane)
Lane 6: K-562 cell lysate (20 µg/Lane)
Lane 7: Human liver tissue lysate (40 µg/Lane)
Lane 8: Mouse liver tissue lysate (40 µg/Lane)
Lane 9: Mouse heart tissue lysate (40 µg/Lane)
Lane 10: Rat liver tissue lysate (40 µg/Lane)
Lane 11: Rat heart tissue lysate (40 µg/Lane)
Lane 12: Zebrafish tissue lysate (40 µg/Lane)
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 5 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 (M1409-2) 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. -
☑ Knockdown (KD)
Western blot analysis of HADH on different lysates with Mouse anti-HADH antibody (M1409-2) 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: 9 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 (M1409-2) 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 (M1409-2) 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 Mouse anti-HADH antibody (M1409-2) at 1/100 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 kidney tissue with Mouse anti-HADH antibody (M1409-2) 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 (M1409-2) 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 colon carcinoma tissue with Mouse anti-HADH antibody (M1409-2) 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 (M1409-2) 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 (M1409-2) 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 (M1409-2) 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 (M1409-2) 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 (M1409-2) 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 (M1409-2) 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 (M1409-2) 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"