Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319)

Overview

  • Product name
    Anti-Lactate Dehydrogenase B antibody [60H11]
    See all Lactate Dehydrogenase B primary antibodies
  • Description
    Mouse monoclonal [60H11] to Lactate Dehydrogenase B
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    His tagged recombinant Human lactate dehydrogenase B protein, purified from E.coli.

  • Positive control
    • WB: HeLa, RD, Ramos, A431, Jurkat, K562 and C6 cell lysates. ICC/IF: HeLa cells. IHC-P: Human breast cancer tissue. Flow Cyt: HeLa cells. IP: HeLa whole cell extract.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR3179006 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

Properties

Applications

Our Abpromise guarantee covers the use of ab85319 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 Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
IP Use at an assay dependent concentration.

Use 2µg.

ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

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 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

  • All lanes : Anti-Lactate Dehydrogenase B antibody [60H11] (ab85319) at 1/2000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : RD cell lysate
    Lane 3 : Ramos cell lysate
    Lane 4 : A431 cell lysate
    Lane 5 : Jurkat cell lysate
    Lane 6 : K562 cell lysate
    Lane 7 : C6 cell lysate


    Predicted band size : 37 kDa
    Observed band size : 37 kDa
  • ICC/IF image of ab85319 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 (ab85319, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • IHC image of ab85319 staining in Human breast cancer formalin fixed paraffin embedded tissue 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 ab85319, 5µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Overlay histogram showing HeLa cells stained with ab85319 (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 (ab85319, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • lactate dehydrogenase B was immunoprecipitated using 0.5mg HeLa whole cell extract, 5µg of Mouse monoclonal to Lactate Dehydrogenase B 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85319.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 37kDa: lactate dehydrogenase B

References

This product has been referenced in:
  • Ding F  et al. Ovariectomy induces a shift in fuel availability and metabolism in the hippocampus of the female transgenic model of familial Alzheimer's. PLoS One 8:e59825 (2013). WB ; Mouse . Read more (PubMed: 23555795) »
  • Billiard J  et al. Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells. Cancer Metab 1:19 (2013). Read more (PubMed: 24280423) »

See all 3 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
3e+006 cells
Specification
mouse embryonic stem cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 10 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse brain)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Mouse brain
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 13 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK cells)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
HEK cells
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 13 2016

Thank you for contacting us.

We are sorry. We have not measured a concentration of this particular antibody.

It is assumed that the concentration of the total protein for LF-MA0336 is 0.5˜2 mg per 1ml considering the band size on QC t...

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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...

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Thanks for your enquiry.

I have requested any further information from the lab regarding testing of ab85319 for reactivity with LDHa.

Also, please note that ab4710 is not an antibody product but instead is a tissue slide. I believe ...

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Thank you for your reply and for sending the image again with your protocol information. There does seem to be a lot of background on the blot, but this can be reduced by further diluting the antibody (we recommend 1:5000-1:10000). Once the background ...

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Thank you for contacting us with your question. I could not find the image attached to your email- could you please resend it? I also have a few questions so that I can better help you: 1) Do you have your original order number ...

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