Overview

  • Product nameAnti-LDHA antibodySee all LDHA primary antibodies ...
  • Description
    Rabbit polyclonal to LDHA
  • Tested applicationsWB, IP, ICC/IF, IHC-P more details
  • Species reactivity
    Reacts with: Human, Zebrafish
    Predicted to work with: Mouse, Rat, Rabbit, Chimpanzee, Chinese Hamster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 to the C-terminus of Human Lactate Dehydrogenase.

    (Peptide available as ab47009.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line)

Properties

Applications

Our Abpromise guarantee covers the use of ab47010 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).Can be blocked with Human Lactate Dehydrogenase peptide (ab47009).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • PathwayFermentation; pyruvate fermentation to lactate; (S)-lactate from pyruvate: step 1/1.
  • Involvement in diseaseDefects 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 similaritiesBelongs to the LDH/MDH superfamily. LDH family.
  • Post-translational
    modifications
    ISGylated.
  • Cellular localizationCytoplasm.
  • Target information above from: UniProt accession P00338 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • Cell proliferation-inducing gene 19 protein antibody
    • GSD11 antibody
    • L lactate dehydrogenase A chain antibody
    • L-lactate dehydrogenase A chain antibody
    • l7R2 antibody
    • Lactate dehydrogenase 1, A chain antibody
    • Lactate dehydrogenase A antibody
    • Lactate dehydrogenase A4 antibody
    • Lactate dehydrogenase M antibody
    • LDH A antibody
    • LDH M antibody
    • LDH muscle subunit antibody
    • LDH muscle subunit; M LDH antibody
    • LDH-A antibody
    • LDH-M antibody
    • LDH1 antibody
    • ldha antibody
    • LDHA_HUMAN antibody
    • LDHM antibody
    • OTTMUSP00000017774 antibody
    • PIG19 antibody
    • Proliferation-inducing gene 19 antibody
    • Renal carcinoma antigen NY-REN-59 antibody
    see all

Anti-LDHA antibody images

  • LDHA was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LDHA and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47010.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 36kDa; LDHA

  • All lanes : LDHA antibody (ab47010) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 37 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab47010 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab47010, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
  • IHC image of Lactate Dehydrogenase staining in human breast carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab47010, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • ab47010 at a 1/400 dilution staining LDHA in human skeletal muscle sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections) incubated for 20 minutes at 25°C. Heat mediated antigen retrieval used, EDTA ph 9.0, 100°C for 20 minutes. Secondary used undiluted, goat anti-mouse/rabbit IgG conjugated to HRP polymer.

    See Abreview


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 37 kDa


    Exposure time : 10 seconds

References for Anti-LDHA antibody (ab47010)

This product has been referenced in:
  • Braunschweig D  et al. Autism-specific maternal autoantibodies recognize critical proteins in developing brain. Transl Psychiatry 3:e277 (2013). Read more (PubMed: 23838888) »
  • Elustondo PA  et al. Physical and functional association of lactate dehydrogenase (LDH) with skeletal muscle mitochondria. J Biol Chem 288:25309-17 (2013). Read more (PubMed: 23873936) »

See all 2 Publications for this product

Product Wall

The antibody ab85319, anti-LDHb, reacts with western blots of recombinant LDHb but not blots containing recombinant LDHa. Please let us know if you have any questions.

As my colleague mentioned in a previous message, ab4710 is not an antibod...

Read More

Thank you for contacting Abcam. I’ve had a look into this and done a bit of a web search and as far as I can see LDH5, LDHM and LDHA are all different names for the same protein, as stated in this paper, http://www.pnas.org/content/107/5/2037.long....

Read More
Application Western blot
Sample Zebrafish Cell lysate - whole cell (zebrafish embryo - 14dpf)
Loading amount 10 µg
Specification zebrafish embryo - 14dpf
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Oct 21 2011

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (skeletal muscle)
Specification skeletal muscle
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA pH 9, 100C, 20 minutes
Permeabilization No
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Submitted Dec 10 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"