Rabbit Recombinant Monoclonal Lipoamide Dehydrogenase antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: PBS
IHC-P | IP | Flow Cyt | WB | ICC/IF | |
---|---|---|---|---|---|
Human | Tested | Not recommended | Not recommended | Tested | Tested |
Mouse | Expected | Not recommended | Not recommended | Tested | Tested |
Rat | Expected | Not recommended | Not recommended | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
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Lipoamide dehydrogenase is a component of the glycine cleavage system as well as an E3 component of three alpha-ketoacid dehydrogenase complexes (pyruvate-, alpha-ketoglutarate-, and branched-chain amino acid-dehydrogenase complex) (PubMed:15712224, PubMed:16442803, PubMed:16770810, PubMed:17404228, PubMed:20160912, PubMed:20385101). The 2-oxoglutarate dehydrogenase complex is mainly active in the mitochondrion (PubMed:29211711). A fraction of the 2-oxoglutarate dehydrogenase complex also localizes in the nucleus and is required for lysine succinylation of histones: associates with KAT2A on chromatin and provides succinyl-CoA to histone succinyltransferase KAT2A (PubMed:29211711). In monomeric form may have additional moonlighting function as serine protease (PubMed:17404228). Involved in the hyperactivation of spermatazoa during capacitation and in the spermatazoal acrosome reaction (By similarity).
GCSL, LAD, PHE3, DLD, Dihydrolipoamide dehydrogenase, Glycine cleavage system L protein
Rabbit Recombinant Monoclonal Lipoamide Dehydrogenase antibody. Carrier free. Suitable for IHC-P, WB, ICC/IF and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: PBS
ab240067 is the carrier-free version of Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Lanes 1-3: Merged signal (red and green). Green - Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 observed at 56 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 Anti-Lipoamide Dehydrogenase antibody [EPR6635] was shown to specifically react with Lipoamide Dehydrogenase in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell line ab265535 (knockout cell lysate Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell lysate ab257922) was used. Wild-type and Lipoamide Dehydrogenase knockout samples were subjected to SDS-PAGE. Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: DLD knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell line (Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell line ab265535)
Lane 3: Jurkat cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 56 kDa
Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Lipoamide Dehydrogenase with Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 antibody at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Lipoamide Dehydrogenase with Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 antibody at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse heart tissue lysate at 20 µg
Lane 3: Mouse kidney tissue lysate at 20 µg
Lane 4: Mouse spleen tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 54 kDa
Exposure time: 1s
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Immunocytochemistry/ Immunofluorescence analysis of C6 (rat glial tumor glial cell) labeling Lipoamide Dehydrogenase with Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
Confocal image showing Mitochondria staining in RAW 264.7 cell line.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) labeling Lipoamide Dehydrogenase with Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
Confocal image showing Mitochondria staining in RAW 264.7 cell line.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte from peripheral blood) labeling Lipoamide Dehydrogenase with Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551 at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at 1/200 was used as the counterstain antibody. Nuclear counter stain was DAPI.
Confocal image showing Mitochondria staining in Jurkat cell line.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (Anti-Lipoamide Dehydrogenase antibody [EPR6635] ab133551) at 1/1000 dilution
Lane 1: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg
Lane 2: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) at 20 µg
Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell) at 20 µg
Lane 4: NIH/3T3 (mouse embryonic fibroblast) at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 54 kDa
Exposure time: 1s
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