Product nameAnti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y]
See all Lactate Dehydrogenase B/LDH-B primary antibodies
DescriptionRabbit monoclonal [EP1565Y] to Lactate Dehydrogenase B/LDH-B
Tested applicationsSuitable for: IHC-Fr, WB, IP, Flow Cyt, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Lactate Dehydrogenase B/LDH-B aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P07195
- WB: Hap1, HEK-293T and HeLa cell lysates. ICC/IF: HepG2. IHC-P: Human heart muscle. Flow Cyt: HeLa cells.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Previously labelled as Lactate Dehydrogenase B.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
Concentration information loading...
PurityProtein A purified
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab53292 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration. PubMed: 22125622|
|WB||1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/100 - 1/250.|
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
Lane 1: Wild-type Hap1 cell lysate (20 µg)
Lane 2: LDHB knockout Hap1 cell lysate (20 µg)
Lane 3: Wild-type HEK-293T cell lysate (20 µg)
Lane 4: LDHB knockout HEK-293T cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab53292 observed at 37 kDa. Red - loading control, ab130007 observed at 125 kDa.
ab53292 was shown to react with Lactate Dehydrogenase in wild-type HEK-293T cells. Loss of signal was observed when knockout sample ab263761 was used. Wild-type and Lactate Dehydrogenase knockout samples were subjected to SDS-PAGE. ab53292 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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 : Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) at 1/50000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : LDHB (Lactate Dehydrogenase B) knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 37 kDa
Lanes 1 - 3: Merged signal (red and green). Green - ab53292 observed at 37 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab53292 was shown to specifically react with Lactate Dehydrogenase B in wild-type HAP1 cells as signal was lost in LDHB (Lactate Dehydrogenase B) knockout cells. Wild-type and LDHB (Lactate Dehydrogenase B) knockout samples were subjected to SDS-PAGE. Ab53292 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab53292 stained HepG2 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 (ab53292, neat) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
Human heart muscle labeled with ab53292 at 1/100 - 1/250 dilution
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) + HeLa cell lysate at 10 µg
Goat anti-rabbit HRP
Predicted band size: 37 kDa
Observed band size: 37 kDa
Overlay histogram showing HeLa cells stained with ab53292 (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 (ab53292, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This product has been referenced in:
- Reynoso E et al. Thioredoxin-1 actively maintains the pseudokinase MLKL in a reduced state to suppress disulfide bond-dependent MLKL polymer formation and necroptosis. J Biol Chem 292:17514-17524 (2017). Read more (PubMed: 28878015) »
- Berrios C et al. Merkel Cell Polyomavirus Small T Antigen Promotes Pro-Glycolytic Metabolic Perturbations Required for Transformation. PLoS Pathog 12:e1006020 (2016). WB ; Human . Read more (PubMed: 27880818) »