Anti-Lactate Dehydrogenase antibody (ab47010)
- Datasheet
- Specific References (14)
- Protocols
Overview
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Product nameAnti-Lactate Dehydrogenase antibody
See all Lactate Dehydrogenase primary antibodies -
DescriptionRabbit polyclonal to Lactate Dehydrogenase
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Host speciesRabbit
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Specificityab47010 cross-reacts with LDHA, B and C.
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Tested applicationsSuitable for: WB, IP, ICC/IF, IHC-Pmore details
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Species reactivityReacts with: Human, Zebrafish
Predicted to work with: Mouse, Rat, Rabbit, Chimpanzee, Chinese hamster
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Immunogen
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Positive control
- This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line)
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Immunohistochemistry kits
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab47010 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).Can be blocked with Human Lactate Dehydrogenase peptide (ab47009). | |
| IP | Use a concentration of 5 µg/ml. | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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PathwayFermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1.
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Involvement in diseaseDefects in LDHA are the cause of glycogen storage disease type 11 (GSD11) [MIM:612933]. A metabolic disorder that results in exertional myoglobinuria, pain, cramps and easy fatigue.
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Sequence similaritiesBelongs to the LDH/MDH superfamily. LDH family.
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Post-translational
modificationsISGylated. -
Cellular localizationCytoplasm.
- Information by UniProt
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Database links
- Entrez Gene: 3939 Human
- Entrez Gene: 3945 Human
- Entrez Gene: 16828 Mouse
- Entrez Gene: 16832 Mouse
- Entrez Gene: 24533 Rat
- Entrez Gene: 24534 Rat
- Entrez Gene: 30496 Zebrafish
- Entrez Gene: 30497 Zebrafish
see all -
Alternative names
- Cell proliferation-inducing gene 19 protein antibody
- GSD11 antibody
- L lactate dehydrogenase B chain antibody
see all
Images
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Western blot - Anti-Lactate Dehydrogenase antibody (ab47010)All lanes : Anti-Lactate Dehydrogenase antibody (ab47010)
Lane 1 : LDHA recombinant
Lane 2 : LDHB recombinant
Lane 3 : LDHC recombinant
Lane 4 : HeLa
Predicted band size: 37 kDa
Exposure time: 30 secondsObserved bands
Lane 1 :40 kDa
Lane 2:37 kDa
Lane 3:62 kDa
Lane 4: 37 kDa
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody (ab47010)IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47010, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase antibody (ab47010)ICC/IF image of ab47010 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47010, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). -
Immunoprecipitation - Anti-Lactate Dehydrogenase antibody (ab47010)LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LDHA and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47010.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 36kDa; LDHA
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Western blot - Anti-Lactate Dehydrogenase antibody (ab47010)All lanes : LDHA antibody (ab47010) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 36 kDa (why is the actual band size different from the predicted?) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody (ab47010)This image is from an anonymous abreview.ab47010 at a 1/400 dilution staining LDHA in human skeletal muscle sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) incubated for 20 minutes at 25°C. Heat mediated antigen retrieval used, EDTA ph 9.0, 100°C for 20 minutes. Secondary used undiluted, goat anti-mouse/rabbit IgG conjugated to HRP polymer. -
Western blot - Anti-Lactate Dehydrogenase antibody (ab47010)This image is courtesy of an anonymous AbreviewAnti-Lactate Dehydrogenase antibody (ab47010) at 1/500 dilution + Zebrafish whole cell lysate at 10 µg
Secondary
HRP conjugated Goat anti-rabbit monoclonal secondary antibody at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 36 kDa (why is the actual band size different from the predicted?)
Additional bands at: 22 kDa (possible non-specific binding)
Exposure time: 30 secondsBlocking step: 5% Milk for 1 hour at 24ºC
Primary antibody incubation for 12 hours at 4ºC
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Western blot - Anti-Lactate Dehydrogenase antibody (ab47010)Anti-Lactate Dehydrogenase antibody (ab47010) at 1 µg/ml +Recombinant human LDHA protein (His tag) (ab93699) at 0.1 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 37 kDa
Exposure time: 10 seconds
Protocols
References
This product has been referenced in:
- Gaude E et al. NADH Shuttling Couples Cytosolic Reductive Carboxylation of Glutamine with Glycolysis in Cells with Mitochondrial Dysfunction. Mol Cell 69:581-593.e7 (2018). Read more (PubMed: 29452638) »
- Olsen RR et al. MYCN induces neuroblastoma in primary neural crest cells. Oncogene 36:5075-5082 (2017). IHC . Read more (PubMed: 28459463) »
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