Anti-Lactate Dehydrogenase antibody (ab47010)

Overview

  • Product name
    Anti-Lactate Dehydrogenase antibody
    See all Lactate Dehydrogenase primary antibodies
  • Description
    Rabbit polyclonal to Lactate Dehydrogenase
  • Specificity
    ab47010 cross-reacts with LDHA, B and C.
  • Tested applications
    Suitable for: WB, IP, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human, Zebrafish
    Predicted to work with: Mouse, Rat, Rabbit, Chimpanzee, Chinese hamster
  • Immunogen

    Synthetic peptide corresponding to Human Lactate Dehydrogenase aa 300 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH).
    (Peptide available as ab47009, 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 Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).
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

Images

  • All lanes : Anti-Lactate Dehydrogenase antibody (ab47010)

    Lane 1 : LDHA recombinant
    Lane 2 : LDHB recombinant
    Lane 3 : LDHC recombinant
    Lane 4 : HeLa


    Predicted band size : 37 kDa


    Exposure time : 30 seconds

    Observed bands

    Lane 1 :40 kDa

    Lane 2:37 kDa

    Lane 3:62 kDa

    Lane 4: 37 kDa

  • 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.
  • 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).
  • 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?)
  • 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

  • Anti-Lactate Dehydrogenase antibody (ab47010) at 1/500 dilution + Zebrafish whole cell lysate at 10 µg

    Secondary
    HRP conjugated Goat anti-rabbit monoclonal secondary antibody at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 37 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 22 kDa (possible non-specific binding).

    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    Blocking step: 5% Milk for 1 hour at 24ºC

    Primary antibody incubation for 12 hours at 4ºC

    See Abreview

  • Anti-Lactate Dehydrogenase antibody (ab47010) at 1 µg/ml + Recombinant human LDHA protein (His tag) (ab93699) at 0.1 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 37 kDa


    Exposure time : 10 seconds

References

This product has been referenced in:
  • Li Y  et al. Mitochondrial pyruvate carrier function is negatively linked to Warburg phenotype in vitro and malignant features in esophageal squamous cell carcinomas. Oncotarget 8:1058-1073 (2017). WB . Read more (PubMed: 27911865) »
  • Li J  et al. Suppression of lactate dehydrogenase A compromises tumor progression by downregulation of the Warburg effect in glioblastoma. Neuroreport 27:110-5 (2016). WB ; Human . Read more (PubMed: 26694942) »

See all 12 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
50 µg
Specification
Heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Dec 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cow Cell lysate - whole cell (VVEC)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
ATP 100 uM TC
Specification
VVEC
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 21 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung)
Antigen retrieval step
None
Permeabilization
No
Specification
lung
Blocking step
peroxidase as blocking agent for 5 minute(s) · Concentration: 0.03%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 18 2015

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Liver)
Permeabilization
No
Specification
Liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Apr 16 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (SH-SY5Y cells)
Specification
SH-SY5Y cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jan 27 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (A 549 cells)
Specification
A 549 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Christine Vohwinkel

Verified customer

Submitted Nov 18 2014

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
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 21 2011

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 (skeletal muscle)
Specification
skeletal muscle
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH 9, 100C, 20 minutes
Permeabilization
No
Username

Abcam user community

Verified customer

Submitted Dec 10 2009

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

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