Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)

Reviews (1)Q&A (1)References (9)
Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • Flow Cytometry - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
  • 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

  • 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.

  • 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
    For more information see here.

    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

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
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 (1)
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.
Notes
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

  • 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

  • Database links

    • Alternative names

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

    Images

    • Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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 kDa



      Lanes 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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.
    • Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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 kDa



      Lanes 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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.

    • Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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

    • Flow Cytometry - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      Flow Cytometry - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      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.

    • Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)
      Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)

    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.

    Click here to view the general protocols

    References (9)

    Publishing research using ab53292? Please let us know so that we can cite the reference in this datasheet.

    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

    Customer reviews and Q&As

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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA Buffer pH 9.0
    Permeabilization
    Yes - Wash Buffer from Dako with Tween
    Specification
    Tonsil
    Fixative
    Paraformaldehyde
    Read More

    Mr. Rudolf Jung

    Verified customer

    Submitted Jul 19 2018

    Question

    To whom it may concern, I am looking for an anti-LDHB antibody. I found the ab53292 antibody on the Epitomics website (Cat.#: 2090-1), which states that it is specific for the B isoform and that it can be bought through abcam. However, this product on the abcam site does not mention any specificity for an isoform. Is it the same one as the Epitomics antibody and with that specific for LDHB? Is there any cross-reactivity with other isoforms? Thank you for your help.

    Read More
    Answer

    Vielen Dank für Ihre Anfrage.

    Es handelt sich hierbei um das selbe Epitomics- Produkt, da wir unsere Produktpalette mit diesen Antikörpern erweitert haben, und diese Antikörper wurden 1:1 aus dem Epitomics Repertoire übernommen, und auch noch im Epitomics Labor hergestellt.

    Das es sich hierbei um den selben Antikörper handelt können sie übrigens auch an der Klon-Nummer erkennen. Da auch das Immunogen das gleiche ist, sollte die Spezifität mit der Isoform B gegeben sein. Und in der Tat kann man diesen Hinweis auf unserem Datenblatt finden:



    A synthetic peptide corresponding to residues near the N-terminus of human LDH-B.


    Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
    https://www.abcam.com/abreviews

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