Recombinant
RabMAb

Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)

Overview

  • Product name
    Anti-Lactate Dehydrogenase antibody [EP1566Y]
    See all Lactate Dehydrogenase primary antibodies
  • Description
    Rabbit monoclonal [EP1566Y] to Lactate Dehydrogenase
  • Specificity
    This antibody reacts with Lactate dehydrogenase
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Lactate Dehydrogenase aa 250 to the C-terminus.

  • Positive control
    • Human liver carcinoma tissue and Hela cell lysate
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
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.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • 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
    modifications
    ISGylated.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • Cell proliferation-inducing gene 19 protein antibody
    • GSD11 antibody
    • L lactate dehydrogenase B chain antibody
    • L-lactate dehydrogenase A chain antibody
    • Lactate dehydrogenase A antibody
    • Lactate dehydrogenase B antibody
    • Lactate dehydrogenase H chain antibody
    • Lactate dehydrogenase M antibody
    • LDH A antibody
    • LDH B antibody
    • LDH H antibody
    • LDH heart subunit antibody
    • LDH M antibody
    • LDH muscle subunit antibody
    • LDH-A antibody
    • LDH-M antibody
    • LDH1 antibody
    • ldha antibody
    • LDHA_HUMAN antibody
    • LDHBD antibody
    • LDHM antibody
    • MS1111 antibody
    • PIG19 antibody
    • Proliferation inducing gene 19 antibody
    • Renal carcinoma antigen NY REN 46 antibody
    • Renal carcinoma antigen NY-REN-59 antibody
    • TRG 5 antibody
    • TRG5 antibody
    see all

Anti-Lactate Dehydrogenase antibody [EP1566Y] images

  • 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.

  • ab52488 immunoprecipitating Lactate Dehydrogenase. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/30 and VeriBlot for IP secondary antibody (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.

  • 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.

  • 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.

  • 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
    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.
  • 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
    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
  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma using unpurified ab52488 at a 1/50 dilution.

  • 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.

  • 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.

References for Anti-Lactate Dehydrogenase antibody [EP1566Y] (ab52488)

This product has been referenced in:
  • Murphy S  et al. Proteomic profiling of mdx-4cv serum reveals highly elevated levels of the inflammation-induced plasma marker haptoglobin in muscular dystrophy. Int J Mol Med 39:1357-1370 (2017). Read more (PubMed: 28440464) »
  • Nassal DM  et al. KChIP2 is a core transcriptional regulator of cardiac excitability. Elife 6:N/A (2017). WB, IF . Read more (PubMed: 28263709) »

See all 24 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12 cells)
Gel Running Conditions
Reduced Denaturing (9%)
Loading amount
30 µg
Treatment
2mM Metformin for 24hr
Specification
C2C12 cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 13 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 18 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 18 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Spinal Cord)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
Spinal Cord
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 26 2015

Application
Western blot
Sample
Human Tissue lysate - whole (Brain Tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
Brain Tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 22 2015

Merci pour votre appel de hier. Je suis désolée du retard de ma réponse.

Voici donc la liste des produits que nous avions discuté ainsi que les liens pour les protocoles que nous avons en ligne.

Les antic...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (pancreas)
Specification
pancreas
Fixative
acetone:methonal (1:1)
Permeabilization
Yes - triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 26 2009

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

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