Anti-ACAA2 antibody [EPR6732(2)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

ab128929, at a 1/50 dilution, staining Human ACAA2 in kidney, using Immunohistochemistry, Formalin/PFA-fixed paraffin-emebedded tissue.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

ab128929, at a 1/250 dilution, staining Human ACAA2 in HepG2 cells, using Immunofluorescence.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

Flow Cytometry analysis of RPMI-8226 (Human plasmacytoma, myeloma B Lymphocyte) cells labeling ACAA2 with purified ab128929 at 1/100 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

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Western blot - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

All lanes:

Western blot - Anti-ACAA2 antibody [EPR6732(2)] (ab128929) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Caco-2 cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 41 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-ACAA2 antibody [EPR6732(2)] (AB128929)

ACAA2 western blot using anti-ACAA2 antibody [EPR6732(2)] ab128929. Publication image and figure legend from Zhang, Y. K., Qu, Y. Y., et al., 2017, Nat Commun, PubMed 28878358.

ab128929 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab128929 please see the product overview.

ECHS1 protein levels are decreased by nutrients. a Schematic diagram of catabolic pathways of FA (black-colored) and BCAA (red-colored) oxidation that involve ECHS1. Long chain FAs are oxidized to short-chained intermediates prior to being completely oxidized through ECHS1. BCAAs are oxidized through ECHS1. ACADL acyl-CoA dehydrogenase, long chain, ACADM medium-chain specific acyl-CoA dehydrogenase, mitochondrial, ACADS short-chain specific acyl-CoA dehydrogenase, ACSL acyl-CoA synthetase long-chain, BCAT branched-chain amino-acid transaminase, BCKDH branched-chain keto acid dehydrogenase, carnitine palmitoyltransferase, ECHS1 enoyl CoA hydratase, short chain 1, mitochondrial, EHHADH Enoyl-CoA, Hydratase/3-Hydroxyacyl CoA Dehydrogenase. b–d Endogenous ECHS1 levels were detected in HEK293T cells treated with different concentrations of glucose (b), FAs (linoleic acid + palmitic acid) (c), and amino acids (glutamate + aspartate) (d). Representative western blot results and quantitation (herein after) of triplicated western blot are shown. e HepG2 cells were exposed to indicated concentrations of glucose. Levels of endogenous β-oxidation enzymes were detected 4 h after glucose exposure. CPT1A Carnitine O-palmitoyltransferase 1, liver isoform, Carnitine O-palmitoyltransferase 2, mitochondrial, HADHA Trifunctional enzyme subunit alpha, mitochondrial, ACAA2 3-ketoacyl-CoA thiolase, mitochondrial. f Plasma ECHS1 levels of normal people (blue, n = 14) and untreated patients with diabetes (red, n = 12) were determined by western blots. Relative ECHS1 levels (normalized to the average ECHS1 level) were plotted against serum glucose levels. g Mitochondria of HEK293T cells cultured in DMEM base, DMEM base supplemented with glucose (25 mM), fatty acids (250 nM) and amino acids (8 mM), respectively, were isolated and the relative (to SOD2 and TOM40) mitochondrial ECHS1 levels were compared. For b–d, mean values of quantitation with SD are reported. *p < 0.05; **p < 0.01; ***p < 0.001

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