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: ICC/IF, WB, IP, Flow Cyt, IHC-P
- Reacts with: Mouse, Rat, Human
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 & Species
Application Species Flow Cyt MouseHumanICC/IF HumanIHC-P MouseHumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide within Human Lactate Dehydrogenase aa 250 to the C-terminus. The exact sequence is proprietary.
Database link: P07195 -
Positive control
- WB: HeLa, HEK293, human heart tissue; rat and mouse kidney tissue lysate. ICC/IF: HeLa cells. Flow cyt: 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.
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Mouse
Human
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ICC/IF |
Human
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IHC-P |
Mouse
Human
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IP |
Human
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WB |
Mouse
Rat
Human
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Application | Abreviews | Notes |
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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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Flow Cyt |
1/50 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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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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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. |
Flow Cyt
1/50 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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) -
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 Fluor® 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)
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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)
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 (38)
ab52488 has been referenced in 38 publications.
- Famulari ES et al. Human liver stem cells express UGT1A1 and improve phenotype of immunocompromised Crigler Najjar syndrome type I mice. Sci Rep 10:887 (2020). PubMed: 31965023
- Pires Da Silva J et al. SIRT1 Protects the Heart from ER Stress-Induced Injury by Promoting eEF2K/eEF2-Dependent Autophagy. Cells 9:N/A (2020). PubMed: 32059483
- Eriksson I et al. Restoration of lysosomal function after damage is accompanied by recycling of lysosomal membrane proteins. Cell Death Dis 11:370 (2020). PubMed: 32409651
- Cardoso HJ et al. Tyrosine kinase inhibitor imatinib modulates the viability and apoptosis of castrate-resistant prostate cancer cells dependently on the glycolytic environment. Life Sci 218:274-283 (2019). PubMed: 30605651
- Le Page LM et al. Assessing the effect of hypoxia on cardiac metabolism using hyperpolarized 13 C magnetic resonance spectroscopy. NMR Biomed 32:e4099 (2019). PubMed: 31090979