The microplate assay ab140361 is used to determine Lactate dehydrogenase B / LDH-B activity (LDHB) in a sample utilizing the immunocapture technique.
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- Cellular and molecular life sciences : CMLS 79:4452022Targeting lactate dehydrogenase B-dependent mitochondrial metabolism affects tumor initiating cells and inhibits tumorigenesis of non-small cell lung cancer by inducing mtDNA damage.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHaibin Deng,Yanyun Gao,Verdiana Trappetti,Damian Hertig,Darya Karatkevich,Tereza Losmanova,Christian Urzi,Huixiang Ge,Gerrit Adriaan Geest,Remy Bruggmann,Valentin Djonov,Jean-Marc Nuoffer,Peter Vermathen,Nicola Zamboni,Carsten Riether,Adrian Ochsenbein,Ren-Wang Peng,Gregor Jan Kocher,Ralph Alexander Schmid,Patrick Dorn,Thomas Michael MartiPubMed 35877003
- Cells 11:2022A Heme-Binding Transcription Factor BACH1 Regulates Lactate Catabolism Suggesting a Combined Therapy for Triple-Negative Breast Cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJoselyn Padilla,Bok-Soon Lee,Karen Zhai,Bethany Rentz,Tia Bobo,Norca Maritza Dowling,Jiyoung LeePubMed 35406740
- Parasitology research 115:1595-6032016Antibodies against Clonorchis sinensis LDH could cross-react with LDHB localizing on the plasma membrane of human hepatocarcinoma cell SMMC-7721 and induce apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTianzhang Song et. al.PubMed 26769711
Key facts
- Detection method
- Colorimetric
- Sample types
- Purified protein, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Adherent cells, Cell Lysate
- Assay type
- Enzyme activity
- Reactive species
- Goat, Cow, Human
- Range
- 8 - 500 ng/mL
Target data
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The microplate assay ab140361 is used to determine Lactate dehydrogenase B / LDH-B activity (LDHB) in a sample utilizing the immunocapture technique.
Key facts
- Detection method
- Colorimetric
- Sample types
- Purified protein, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Adherent cells, Cell Lysate
- Assay type
- Enzyme activity
- Reactive species
- Goat, Cow, Human
- Range
- 8 - 500 ng/mL
- Assay Platform
- Microplate reader
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 3 | C.V. 5 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Overall | n 4 | C.V. 5 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 95 | Range 86 - 111 % |
Sample type Cell culture media | Average % = 94 | Range 91 - 101 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- Multi
- Appropriate long-term storage conditions
- Multi
- Storage information
- Please refer to protocols
Notes
The microplate assay ab140361 is used to determine Lactate dehydrogenase B / LDH-B activity (LDHB) in a sample utilizing the immunocapture technique. The specific native LDHB enzyme is immune-captured within the wells of the microplate and activity is determined by following the production of NADH catalyzed by the enzyme. The generation of NADH is coupled to the 1:1 reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm (Dye molar extinction coefficient - 37000 M-1 cm-1). The immune-capture based activity assay kit contains all the required reagents for a fast and simple measurement of LDHB activity in a whole cell extract, tissue homogenate, or serum.
Functional lactate dehydrogenase are homo or hetero tetramers composed of M and H protein subunits encoded by the LDHA and LDHB genes, respectively. The major isozymes of skeletal muscle and liver has four muscle (M) subunits; while the main isozymes for heart muscle contains four (H) subunits. The other variants contain both types of subunits. Elevated levels of LDH indicate tissue breakdown in hemolysis. LDH can be used as a marker of myocardial infarction, peaking at days 3 and lasting through day 10. Other disorders indicated by elevated LDH include cancer, meningitis, encephalitis, acute pancreatitis, and HIV. Defects in LDHB are a cause of hereditary LDHB deficiency; however LDHB deficiency is usually asymptomatic.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Lactate Dehydrogenase B (LDH-B) also known as LDH is an enzyme that plays an important role in the interconversion of lactate to pyruvate. It belongs to the lactate dehydrogenase family which includes multiple isoenzymes. LDH-B has a molecular weight of approximately 36 kDa and commonly forms a tetramer with three other similar subunits. It is mainly expressed in heart and muscle tissues among others where it functions effectively.
- Biological function summary
LDH-B is essential for cellular energy production processes. It catalyzes the conversion of pyruvate a byproduct of glycolysis into lactate while simultaneously converting NADH to NAD+. This reaction is significant under anaerobic conditions where mitochondria-based ATP production is limited. LDH-B does not function alone and can operate as part of the lactate dehydrogenase complex containing different subunit combinations.
- Pathways
Lactate and pyruvate metabolism engage LDH-B facilitating the Cori cycle and the lactate shuttle. These pathways enable lactate transport between tissues such as muscle and liver and they are important in maintaining blood glucose levels and energy balance. LDH-B operates alongside its related isoenzymes such as LDH-A which predominantly act in specific tissues and conditions to optimize energy utilization.
- Associated diseases and disorders
Anomalous LDH-B activity associates with conditions like myocardial infarction and some cancer types. Enhanced expression or activity of LDH-B can indicate altered metabolic states in tumors connecting it as a potential marker for certain cancerous conditions. The relationship with other proteins such as the hypoxia-inducible factor (HIF) is notable especially in cancer where HIF can regulate LDH-B expression under low oxygen environments.
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4 product images
Functional Studies - Lactate Dehydrogenase B / LDH-B Activity Assay (ab140361)
Example control standard curve generaated using the method described in the protocl booklet.
Functional Studies - Lactate Dehydrogenase B / LDH-B Activity Assay (ab140361)
Lysate example curve. LDHB activity was measured from Hela cell lysate, which contains hetero-tetramer isozyme composed of both (M) LDHA and (H) LDHB subunits. However, the liver derived cell line, HepG2, which contains mainly homo-tetramer of subunit M (LDHA), shows no enzymatic activity.
Functional Studies - Lactate Dehydrogenase B / LDH-B Activity Assay (ab140361)
Species cross reactivity shown with sera. The species cross reactivity of this assay was demonstrated by testing the sera from multiple species. The kit shows a good reaction with human, goat and bovine samples, but shows no reaction with mouse or rat sera which may also contain LDHB. This species cross reaction specificity is due to the species specificity of the capture antibody.
Western blot - Lactate Dehydrogenase B / LDH-B Activity Assay (ab140361)
Isoform specificity validated by Western blot.
Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: LDHA (purified from rabbit skeletal muscle)
Lane 4: LDHB (recombinant)
Lane 5: HeLa – In-well extracted with capture mAb
Lane 6: HepG2 – In-well extracted with capture mAb
Lane 7: LDHA – In-well extracted with a capture mAb
Lane 8: LDHB – In-well extracted with capture mAbThe western blot results indicate that the capture antibody isolates active LDHB, but does not isolate LDHA (which would be detected by the anti-LDH antibody).
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