Recombinant Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1565Y] to Lactate Dehydrogenase B/LDH-B
- Suitable for: WB, Flow Cyt, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y]
See all Lactate Dehydrogenase B/LDH-B primary antibodies -
Description
Rabbit monoclonal [EP1565Y] to Lactate Dehydrogenase B/LDH-B -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Lactate Dehydrogenase B/LDH-B aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P07195 -
Positive control
- WB: Hap1, HEK-293T and HeLa cell lysates. ICC/IF: HepG2. IHC-P: Human heart muscle. Flow Cyt: HeLa cells.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1565Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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WB |
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
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Flow Cyt |
1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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Notes |
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WB
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). |
Flow Cyt
1/70. 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. |
Target
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Pathway
Fermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1. -
Involvement in disease
Note=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 similarities
Belongs to the LDH/MDH superfamily. LDH family. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 3945 Human
- Omim: 150100 Human
- SwissProt: P07195 Human
- Unigene: 446149 Human
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Alternative names
- Epididymis secretory protein Li 281 antibody
- HEL S 281 antibody
- L lactate dehydrogenase B chain antibody
see all
Images
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All lanes : Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : LDHB knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDaLanes 1- 2: Merged signal (red and green). Green - ab53292 observed at 37 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
ab53292 was shown to react with Lactate Dehydrogenase B/LDH-B in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255403 (knockout cell lysate ab263761) was used. Wild-type HEK-293T and LDHB knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab53292 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 5000 dilution and a 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.
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Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)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.
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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 kDaLanes 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
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.
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Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) + HeLa cell lysate at 10 µg
Secondary
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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab53292 has been referenced in 9 publications.
- Tasdogan A et al. Metabolic heterogeneity confers differences in melanoma metastatic potential. Nature 577:115-120 (2020). PubMed: 31853067
- 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). 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 . PubMed: 27880818
- Vaughan RA et al. ß-alanine suppresses malignant breast epithelial cell aggressiveness through alterations in metabolism and cellular acidity in vitro. Mol Cancer 13:14 (2014). WB ; Human . PubMed: 24460609
- McCleland ML et al. Lactate dehydrogenase B is required for the growth of KRAS-dependent lung adenocarcinomas. Clin Cancer Res 19:773-84 (2013). WB ; Human . PubMed: 23224736
- Lichtenfels R et al. A proteomic view at T cell costimulation. PLoS One 7:e32994 (2012). Flow Cyt ; Human . PubMed: 22539942
- McCleland ML et al. An integrated genomic screen identifies LDHB as an essential gene for triple-negative breast cancer. Cancer Res 72:5812-23 (2012). Immunomicroscopy ; Human . PubMed: 23139210
- O'Dwyer D et al. The proteomics of colorectal cancer: identification of a protein signature associated with prognosis. PLoS One 6:e27718 (2011). IHC-Fr ; Human . PubMed: 22125622
- Liao AC et al. Loss of lactate dehydrogenase B subunit expression is correlated with tumour progression and independently predicts inferior disease-specific survival in urinary bladder urothelial carcinoma. Pathology 43:707-12 (2011). IHC ; Human . PubMed: 22027740