Recombinant
RabMAb

Recombinant 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
  • Host species
    Rabbit
  • Specificity

    This antibody reacts with Lactate dehydrogenase; LDHA (79%), LDHB (100%) and LDHC (86%).

  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Lactate Dehydrogenase aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: P07195

  • 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 IHC antigen retrieval protocols.

Target

Images

  • 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
    All lanes : 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.

  • 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 Detection Reagent (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 1/20000 dilution (purified)

    Lane 1 : Mouse kidney tissue lysate
    Lane 2 : Rat kidney tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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

This product has been referenced in:
  • Cardoso HJ  et al. Tyrosine kinase inhibitor imatinib modulates the viability and apoptosis of castrate-resistant prostate cancer cells dependently on the glycolytic environment. Life Sci 218:274-283 (2019). Read more (PubMed: 30605651) »
  • Song ES  et al. Insulin-degrading enzyme is not secreted from cultured cells. Sci Rep 8:2335 (2018). Read more (PubMed: 29402917) »
See all 32 Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Lung adenocarcinoma)
Permeabilization
No
Specification
Lung adenocarcinoma
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 8% · Temperature: 27°C
Fixative
Paraformaldehyde

Dr. Shubhangi Agarwal

Verified customer

Submitted Jan 31 2018

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

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

Abcam user community

Verified customer

Submitted Apr 18 2016

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

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

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

Abcam user community

Verified customer

Submitted Jun 22 2015

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

Abcam user community

Verified customer

Submitted Oct 26 2009

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