ETFA Polyclonal antibody

ETFA Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

5

Cat no : 12262-1-AP

Synonyms

MADD, GA2, EMA, Alpha-ETF, Alpha ETF

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Validation Data Gallery

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  WB analysis using 12262-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 12262-1-AP (ETFA antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse small intestine using 12262-1-AP

  mouse small intestine tissue were subjected to SDS PAGE followed by western blot with 12262-1-AP (ETFA antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of human ileum using 12262-1-AP

  human ileum tissue were subjected to SDS PAGE followed by western blot with 12262-1-AP (ETFA antibody) at dilution of 1:400 incubated at room temperature for 1.5 hours.

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  WB analysis of Y79 using 12262-1-AP

  Y79 cells were subjected to SDS PAGE followed by western blot with 12262-1-AP (ETFA antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 12262-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12262-1-AP (ETFA antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 12262-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 12262-1-AP (ETFA antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 12262-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ETFA antibody (12262-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), 594phalloidin (red).

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  FC experiment of MCF-7 using 12262-1-AP

  1x10^6 MCF-7 cells were intracellularly stained with 0.8 ug ETFA Polyclonal antibody (12262-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.8 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Y79 cells, human ileum tissue, mouse small intestine tissue, rat small intestine tissue
• Positive IHC detected in: human breast cancer 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: HeLa cells
• Positive FC (Intra) detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:3000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension

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

• WB: See 4 publications below
• IF: See 1 publications below

Product Information

12262-1-AP targets ETFA in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: ETFA fusion protein Ag2931
• Full Name: electron-transfer-flavoprotein, alpha polypeptide
• Calculated Molecular Weight: 30 kDa
• Observed Molecular Weight: 30 kDa
• GenBank Accession Number: BC015526
• Gene Symbol: ETFA
• Gene ID (NCBI): 2108
• RRID: AB_2100540
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• 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

Electron transfer flavoprotein (ETF) exists in the mitochondrial matrix as a heterodimer of 30 kD alpha subunits (ETFA) and 28 kD beta subunits (ETFB) and contains 1 flavin adenine dinucleotide (FAD) and 1 adenosine 5-prime monophosphate (AMP) per heterodimer. ETFA serves as a specific electron acceptor for several dehydrogenases, including five acyl-CoA dehydrogenases, glutaryl-CoA and sarcosine dehydrogenase.

Publications

Species	Application	Title
human	IF	Mol Cell; Filamentous GLS1 promotes ROS-induced apoptosis upon glutamine deprivation via insufficient asparagine synthesis.; Authors - Bin Jiang
mouse	WB	Cell Mol Life Sci; Mitochondrial FAD shortage in SLC25A32 deficiency affects folate-mediated one-carbon metabolism.; Authors - Min-Zhi Peng
human	WB	J Proteome Res; Proteomic analysis of nuclei isolated from cancer cell lines treated with indenoisoquinoline NSC 724998, a novel topoisomerase I inhibitor.; Authors - Han Bingnan B
human	WB	J Proteomics; Global expression study in colorectal cancer on proteins with alkaline isoelectric point by two-dimensional difference gel electrophoresis.; Authors - Muto Taika T
human	WB	Proteome Sci; Activity-based protein profiling of the hepatitis C virus replication in Huh-7 hepatoma cells using a non-directed active site probe.; Authors - Singaravelu Ragunath R