HADHA Monoclonal antibody

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  WB analysis of HEK293 cells using 60250-1-Ig

  WB result of HADHA specific antibody (60250-1-Ig, 1:4000) with si-control and si-HADHA transfected HEK293 cells.

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  WB analysis using 60250-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60250-1-Ig (HADHA antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 60250-1-Ig

  HepG2 cells were subjected to SDS PAGE followed by western blot with 60250-1-Ig (HADHA Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of Jurkat using 60250-1-Ig

  Jurkat cells were subjected to SDS PAGE followed by western blot with 60250-1-Ig (HADHA Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of mouse heart using 60250-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 60250-1-Ig (HADHA antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse heart using 60250-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse heart tissue slide using 60250-1-Ig (HADHA antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 60250-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using 60250-1-Ig (HADHA antibody) at dilution of 1:100 and CoraLite488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  IF Staining of HepG2 using 60250-1-Ig

  Immunofluorescent analysis of (10% Formaldehyde ) fixed HepG2 cells using 60250-1-Ig(HADHA antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  Affinity and Kinetic Characterization of 60250-1-Ig

  Biolayer interferometry (BLl) kinetic assay of 60250-1-Ig against Human HADHA was performed. The affinity constant is 17.3 nM.

• Featured Product
• KD/KO Validated

HADHA Monoclonal Antibody for WB, IHC, IF/ICC, ELISA

Cat No. 60250-1-Ig

Clone No.3E9B1

Publications(9)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

3E9B1, 78 kDa gastrin binding protein, 78 kDa gastrin-binding protein, EC:2.3.1.-, ECHA

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Tested Applications

• Positive WB detected in: HeLa cells, Jurkat cells, HepG2 cells, HEK293 cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, RAW 264.7 cells
• Positive IHC detected in: mouse heart tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 9 publications below

Product Information

60250-1-Ig targets HADHA in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag1211

  Product name: Recombinant human HADHA protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 339-763 aa of BC009235

  Sequence: LMGLYHGQVLCKKNKFGAPQKDVKHLAILGAGLMGAGIAQVSVDKGLKTILKDATLTALDRGQQQVFKGLNDKVKKKALTSFERDSIFSNLTGQLDYQGFEKADMVIEAVFEDLSLKHRVLKEVEAVIPDHCIFASNTSALPISEIAAVSKRPEKVIGMHYFSPVDKMQLLEIITTEKTSKDTSASAVAVGLKQGKVIIVVKDGPGFYTTRCLAPMMSEVIRILQEGVDPKKLDSLTTSFGFPVGAATLVDEVGVDVAKHVAEDLGKVFGERFGGGNPELLTQMVSKGFLGRKSGKGFYIYQEGVKRKDLNSDMDSILASLKLPPKSEVSSDEDIQFRLVTRFVNEAVMCLQEGILATPAEGDIGAVFGLGFPPCLGGPFRFVDLYGAQKIVDRLKKYEAAYGKQFTPCQLLADHANSPNKKFYQ

• Full Name: hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit
• Calculated Molecular Weight: 83 kDa
• Observed Molecular Weight: 79 kDa
• GenBank Accession Number: BC009235
• Gene Symbol: HADHA
• Gene ID (NCBI): 3030
• RRID: AB_2881371
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: P40939
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

HADHA(Trifunctional enzyme subunit alpha, mitochondrial) is also named as HADH,78 kDa gastrin-binding protein.It belongs to the enoyl-CoA hydratase/isomerase family in the N-terminal section and the 3-hydroxyacyl-CoA dehydrogenase family in the central section.It harbors the 3-hydroxyacyl-CoA dehydrogenase and enoyl-CoA hydratase activities.Defects in HADHA are a cause of trifunctional protein deficiency (TFP deficiency) and long-chain 3-hydroxyl-CoA dehydrogenase deficiency (LCHAD deficiency) and maternal acute fatty liver of pregnancy (AFLP).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	WB	Commun Biol; A G1528C Hadha knock-in mouse model recapitulates aspects of human clinical phenotypes for long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency; Authors - Garen Gaston
human	WB	Front Oncol; Multi-Omics Analysis of Fatty Acid Metabolism in Thyroid Carcinoma.; Authors - Jinghui Lu
mouse	WB	Proteomics; Quantitative Proteomic Study of Myocardial Mitochondria in Urea Transporter B Knockout Mice.; Authors - Yanwei Du ; KO Validated
human	WB	Cell Biosci; Exosomes from hyperglycemia-stimulated vascular endothelial cells contain versican that regulate calcification/senescence in vascular smooth muscle cells.; Authors - Shuang Li
human	WB	JCI Insight; Mitochondrial bioenergetics and cardiolipin remodeling abnormalities in mitochondrial trifunctional protein deficiency; Authors - Eduardo Vieira Neto
human	WB	J Cancer; Knockdown of HOXD13 in Oral Squamous Cell Carcinoma Inhibited its Proliferation, Migration, and Influenced Fatty Acid Metabolism; Authors - Xingyue Ma