Recombinant Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1566Y] to Lactate Dehydrogenase
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Lactate Dehydrogenase antibody [EP1566Y]
See all Lactate Dehydrogenase primary antibodies -
Description
Rabbit monoclonal [EP1566Y] to Lactate Dehydrogenase -
Host species
Rabbit -
Specificity
This antibody reacts with Lactate dehydrogenase; LDHA (79%), LDHB (100%) and LDHC (86%).
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK293, human heart tissue; rat and mouse kidney tissue lysate. ICC/IF: HeLa cells. Flow Cyt (intra): Raw264.7 cells, HeLa. IP: HeLa cells. IHC-P: Mouse liver tissue, human breast carcinoma tissue, human liver carcinoma.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1566Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52488 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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Flow Cyt (Intra) |
1/50 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
1/50 - 1/100.
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WB | (3) |
1/5000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
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IP |
1/30 - 1/100.
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IHC-P | (2) |
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/50 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/50 - 1/100. |
WB
1/5000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). |
IP
1/30 - 1/100. |
IHC-P
1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Pathway
Fermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1. -
Involvement in disease
Defects 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. -
Sequence similarities
Belongs to the LDH/MDH superfamily. LDH family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm. - 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
- Omim: 150000 Human
- Omim: 150100 Human
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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All lanes : Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/5000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK293 (human embryonic kidney) whole cell lysate
Lane 3 : Human heart tissue
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 37 kDa
Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands. -
ab52488 staining Lactate Dehydrogenase in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab52488 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.
Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) -
Intracellular Flow Cytometry analysis of Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate labelling Lactate Dehydrogenase with purified ab52488 at 1/190 (red). Cells were fixed with 4% paraformaldehyde. Alexa Fluorr® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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ab52488 immunoprecipitating Lactate Dehydrogenase. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/30 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.
Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab52488 in HeLa (human cervix adenocarcinoma) whole cell lysate -
ICC/IF image of unpurified ab52488 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 (ab52488, 1µg/ml) overnight at +4°C. 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) 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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ab52488 staining Lactate Dehydrogenase in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
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ab52488 staining Lactate Dehydrogenase in human breast carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. ab97051 was used as the secondary antibody.
Negative control 1: PBS in place of primary antibody.
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All lanes : Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/20000 dilution (purified)
Lane 1 : Mouse kidney tissue lysate
Lane 2 : Rat kidney tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer 5% NFDM/TBST -
Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488) at 1/100000 dilution (unpurified) + Hela cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa -
Immunohistochemical analysis of paraffin-embedded human liver carcinoma using unpurified ab52488 at a 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab52488 staining Lactate Dehydrogenase in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/2000. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.
Negative control 1: PBS in place of primary antibody.
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Overlay histogram showing HeLa cells stained with unpurified ab52488 (red line). The cells were fixed with 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 (ab52488, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a slightly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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 (64)
ab52488 has been referenced in 64 publications.
- Shima T et al. Glucose transporter‑1 inhibition overcomes imatinib resistance in gastrointestinal stromal tumor cells. Oncol Rep 47:N/A (2022). PubMed: 34738628
- Wu F et al. TGF-βRII regulates glucose metabolism in oral cancer-associated fibroblasts via promoting PKM2 nuclear translocation. Cell Death Discov 8:3 (2022). PubMed: 35013150
- Liu Y et al. RIG-I Promotes Cell Viability, Colony Formation, and Glucose Metabolism and Inhibits Cell Apoptosis in Colorectal Cancer by NF-κB Signaling Pathway. Dis Markers 2022:1247007 (2022). PubMed: 35242239
- Jiang Y et al. CircRBM33 downregulation inhibits hypoxia-induced glycolysis and promotes apoptosis of breast cancer cells via a microRNA-542-3p/HIF-1α axis. Cell Death Discov 8:126 (2022). PubMed: 35318311
- Ma H et al. Single-Cell RNA-Sequencing Analyses Revealed Heterogeneity and Dynamic Changes of Metabolic Pathways in Astrocytes at the Acute Phase of Ischemic Stroke. Oxid Med Cell Longev 2022:1817721 (2022). PubMed: 35535357