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    products/assay-kits/lactate-dehydrogenase-b--ldh-b-activity-assay-ab140361.pdf

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Signal Transduction Metabolism Energy Metabolism
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Lactate Dehydrogenase B / LDH-B Activity Assay (ab140361)

  • Datasheet
  • SDS
  • Protocol Booklet
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Example control standard curve
  • Lysate example curve
  • Species cross reactivity shown with sera
  • Isoform specificity validated by Western blot

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: Adherent cells, Cell culture supernatant, Cell Lysate, Plasma, Purified protein, Serum, Tissue Extracts

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Overview

  • Product name

    Lactate Dehydrogenase B / LDH-B Activity Assay
    See all Lactate Dehydrogenase B/LDH-B kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 3 5%
    Inter-assay
    Sample n Mean SD CV%
    Overall 4 5%
  • Sample type

    Cell culture supernatant, Serum, Plasma, Adherent cells, Tissue Extracts, Cell Lysate, Purified protein
  • Assay type

    Enzyme activity
  • Range

    8 ng/ml - 500 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 95 86% - 111%
    Cell culture media 94 91% - 101%
  • Species reactivity

    Reacts with: Goat, Cow, Human
    Does not react with: Mouse, Rat
  • Product overview

    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.

  • Notes

    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.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Coupler 1 unit
    100X NAD+ 1 unit
    100X Reagent Dye 1 unit
    10X Blocking Buffer 1 x 6ml
    10X Buffer 1 x 25ml
    1X LDH-B Control 1 x 1µg
    Extraction Buffer (ab260490) 1 x 15ml
    LDH-B Microplate 1 unit
    Reaction Base Buffer 1 x 10ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Dehydrogenase Kits
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Oxidative Stress Assay Kits
    • NADP/NADPH
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
  • 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.
  • Target information above from: UniProt accession P07195 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Epididymis secretory protein Li 281
    • HEL S 281
    • L lactate dehydrogenase B chain
    • L-lactate dehydrogenase B chain
    • Lactate Dehydrogenase B
    • Lactate dehydrogenase H chain
    • LDH B
    • LDH H
    • LDH heart subunit
    • LDH-B
    • LDH-H
    • LDHB
    • LDHB_HUMAN
    • LDHBD
    • LDHH
    • Renal carcinoma antigen NY REN 46
    • Renal carcinoma antigen NY-REN-46
    • TRG-5
    • TRG5
    see all
  • Database links

    • Entrez Gene: 281275 Cow
    • Entrez Gene: 3945 Human
    • Omim: 150100 Human
    • SwissProt: Q5E9B1 Cow
    • SwissProt: P07195 Human
    • Unigene: 446149 Human

    Associated products

    • Positive Controls

      • Bovine Heart Mitochondria (ab110338)

    Images

    • Example control standard curve
      Example control standard curve

      Example control standard curve generaated using the method described in the protocl booklet.

    • Lysate example curve
      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.
    • Species cross reactivity shown with sera
      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.
    • Isoform specificity validated by Western blot
      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 mAb

      The 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).

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab140361? Please let us know so that we can cite the reference in this datasheet.

    ab140361 has been referenced in 2 publications.

    • Deng H  et al. Targeting lactate dehydrogenase B-dependent mitochondrial metabolism affects tumor initiating cells and inhibits tumorigenesis of non-small cell lung cancer by inducing mtDNA damage. Cell Mol Life Sci 79:445 (2022). PubMed: 35877003
    • Song T  et al. Antibodies against Clonorchis sinensis LDH could cross-react with LDHB localizing on the plasma membrane of human hepatocarcinoma cell SMMC-7721 and induce apoptosis. Parasitol Res 115:1595-603 (2016). PubMed: 26769711

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