Product nameAnti-Lactate Dehydrogenase B/LDH-B antibody [60H11]
See all Lactate Dehydrogenase B/LDH-B primary antibodies
DescriptionMouse monoclonal [60H11] to Lactate Dehydrogenase B/LDH-B
Tested applicationsSuitable for: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant full length protein corresponding to Human Lactate Dehydrogenase B/LDH-B.
- WB: HeLa, RD, Ramos, A431, Jurkat, K562 and C6 cell lysates. ICC/IF: HeLa cells. IHC-P: Human breast cancer tissue. Flow Cyt: HeLa cells. IP: HeLa whole cell extract.
This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR3179006 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
Previously labelled as Lactate Dehydrogenase B.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.03% Sodium azide
Constituents: 0.01% BSA, 50% Glycerol, 0.87% Sodium chloride, HEPES
PurityTissue culture supernatant
Light chain typelambda
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab85319 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).|
|IP||Use at an assay dependent concentration.
|ELISA||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
PathwayFermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1.
Involvement in diseaseNote=Defects in LDHB result in deficiency of lactate dehydrogenase, a condition with no clear symptomatic consequences. Lactate dehydrogenase deficiency can probably be considered a non-disease.
Sequence similaritiesBelongs to the LDH/MDH superfamily. LDH family.
- Information by UniProt
- Epididymis secretory protein Li 281 antibody
- HEL S 281 antibody
- L lactate dehydrogenase B chain antibody
All lanes : Anti-Lactate Dehydrogenase B/LDH-B antibody [60H11] (ab85319) at 1/2000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : RD cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : A431 cell lysate
Lane 5 : Jurkat cell lysate
Lane 6 : K562 cell lysate
Lane 7 : C6 cell lysate
Predicted band size: 37 kDa
Observed band size: 37 kDa
ICC/IF image of ab85319 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85319, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab85319 staining in Human breast cancer formalin fixed paraffin embedded tissue 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 ab85319, 5µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Overlay histogram showing HeLa cells stained with ab85319 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab85319, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Lactate Dehydrogenase B/LDH-B was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Mouse monoclonal to Lactate Dehydrogenase B 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85319.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 37kDa: lactate dehydrogenase B
This product has been referenced in:
- Shi L et al. SIRT5-mediated deacetylation of LDHB promotes autophagy and tumorigenesis in colorectal cancer. Mol Oncol 13:358-375 (2019). Read more (PubMed: 30443978) »
- Choi YS et al. Hyperpolarized [1-13C] pyruvate MR spectroscopy detect altered glycolysis in the brain of a cognitively impaired mouse model fed high-fat diet. Mol Brain 11:74 (2018). Read more (PubMed: 30563553) »