Anti-Monocarboxylic acid transporter 1 antibody (ab85021)

Reviews (4)Q&A (2)References (1)

Overview

  • Product name
    Anti-Monocarboxylic acid transporter 1 antibody
    See all Monocarboxylic acid transporter 1 primary antibodies
  • Description
    Rabbit polyclonal to Monocarboxylic acid transporter 1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human Monocarboxylic acid transporter 1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab97985.)

  • Positive control
    • This antibody gave a positive signal in Human Skeletal Muscle tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab85021 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 54 kDa (predicted molecular weight: 54 kDa).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function
    Proton-linked monocarboxylate transporter. Catalyzes the rapid transport across the plasma membrane of many monocarboxylates such as lactate, pyruvate, branched-chain oxo acids derived from leucine, valine and isoleucine, and the ketone bodies acetoacetate, beta-hydroxybutyrate and acetate.
  • Tissue specificity
    Widely expressed in normal and in cancer cells.
  • Involvement in disease
    Symptomatic deficiency in lactate transport
    Familial hyperinsulinemic hypoglycemia 7
  • Sequence similarities
    Belongs to the major facilitator superfamily. Monocarboxylate porter (TC 2.A.1.13) family.
  • Cellular localization
    Cell membrane.
  • Information by UniProt
  • Database links
    • Alternative names
      • FLJ36745 antibody
      • HHF7 antibody
      • MCT 1 antibody
      • MCT antibody
      • MGC44475 antibody
      • Monocarboxylate transporter 1 antibody
      • Monocarboxylate transporter antibody
      • Monocarboxylate transporter isoform 1 antibody
      • Monocarboxylic acid transporter 1 antibody
      • MOT1_HUMAN antibody
      • Slc16a1 antibody
      • SLC16A1 protein antibody
      • Solute carrier family 16 (monocarboxylic acid transporters) member 1 antibody
      • Solute carrier family 16 member 1 (monocarboxylic acid transporter 1) antibody
      • Solute carrier family 16 member 1 antibody
      see all

    Images

    • Western blot - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      Western blot - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      Anti-Monocarboxylic acid transporter 1 antibody (ab85021) at 1 µg/ml + Human skeletal muscle tissue lysate - total protein (ab29330) at 10 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 54 kDa
      Observed band size: 54 kDa
      Additional bands at: 40 kDa, 89 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 2 minutes
    • Immunocytochemistry/ Immunofluorescence - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      Immunocytochemistry/ Immunofluorescence - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      ICC/IF image of ab85021 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab85021, 5µg/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. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Monocarboxylic acid transporter 1 antibody (ab85021)
      IHC image of Monocarboxylic acid transporter 1 staining in human normal kidney 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 ab85021, 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.

    References

    This product has been referenced in:
    • Jensen DH  et al. A reverse Warburg metabolism in oral squamous cell carcinoma is not dependent upon myofibroblasts. J Oral Pathol Med N/A:N/A (2014). IHC-P ; Human . Read more (PubMed: 25420473) »
    See 1 Publication for this product

    Publishing research using ab85021? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Monocarboxylic acid transporter 1 antibody

     Excellent
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Oral squamous cell carcinoma)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    Oral squamous cell carcinoma
    Blocking step
    VENTANA Benchmark ultra as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 00% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Read More

    David Hebbelstrup Jensen

    Verified customer

    Submitted Jun 29 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Monocarboxylic acid transporter 1 antibody

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
    Sample
    Human Cell (H1299 Lung cancer cells)
    Specification
    H1299 Lung cancer cells
    Permeabilization
    Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
    Fixative
    Paraformaldehyde
    Read More

    Dr. Dimitra Kalamida

    Verified customer

    Submitted Feb 27 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Monocarboxylic acid transporter 1 antibody

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate, pH6, 100C, 20 minutes
    Sample
    Human Tissue sections (appendix testis)
    Specification
    appendix testis
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 15 2014

    Question

    first of all, thanks again for the peptide and my apologies for the
    long time it took me to answer. My PI gave me some urgent tasks, so I
    could do the WB only last week.
    Anyway, the results are quite clear: None of the lower bands
    disappeared, so I can not use them for analysis as binding is
    unspecific (see attachment). However, I modified the protocol one last
    time and incubated both primary and secondary antibody on ice, which
    yielded far less background than extensive blocking and washing steps.
    Therefore, I think I can use the ˜54 kDa band, which is great!
    Again, thank you for the support and time!
    Kind regards,

    Read More
    Answer

    Thank you for your reply.

    I am very happy to hear that the antibody is now also working well in WB.

    We would like to make the improved protocol available to other users and therefore I would like to ask you to write an Abreview for WB ( and if you would like also the other applications that you used ab85021 in).

    This would help other scientist with their protocol and you would earn Abpoints (images always get you 100 extra Abpoints!)that can be redeemed as discount on future purchases or Amazon vouchers.

    We appreciate your feedback very much and wish you good luck with your research.

    Read More

    Question

    thank you very much again for providing a free replacement of MCT1
    antibody ab85021! I tried it now in flow cytometry, IHC and ICC with
    quite satisfactory results. However, in Western Blot the antibody
    still does not perform well. I tried really hard in several blots to
    get rid of the background, but was not really successful. I went up to
    10% BSA and 0.5% Tween 20 as blocking reagent, blocked over night
    (4°C), incubated the antibody in blocking solution (1 h RT) and washed
    in several steps for a total time of more than 4 h after seconary
    antibody incubation. I also tried both nitrocellulose and PVDF
    membrane. Still, the blots are background-heavy (see attachment). When
    I used the secondary antibody with other primary antibodies, there was
    no background at all, so this cannot be the problem.
    As you can see, the expected band at 54 kDa is very faint and embedded
    in a lot of background, so it is not usable at all. However, it is
    frequently published that MCT1 appears at 44 kDa. The topmost band of
    the three distict bands (in the red box) might be something like 41-42
    kDa and might therefore be usable. When discussing this issue with my
    PI, she said this might be proven if we mix the antibody with the
    immunizing peptide. As the results for the other methods look OK and I
    do not want to claim that the antibody is not working and ask for a
    replacement yet again (and in addition, the blot shown in the Abreview
    section does not resemble the data sheet, neither), my suggestion
    would be to compare +/- immunizing peptide and see what bands
    dissappear.
    I would really appreciate your opinion on that subject and how we
    should continue from hereon.
    Best regards,

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. I contacted our in house QC department and they will have a closer look at ab85021.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free blocking peptide (ab97985) with the order number 1134611.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I would like to aks you for your feedback after using this blocking peptide. this information is very valuable for the QC department. Thank you.

    I wish you the best of luck with your research.

    Read More

    Western blot abreview for Anti-Monocarboxylic acid transporter 1 antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (glioma cell, placenta, MDCK, HEK, HL-1)
    Loading amount
    35 µg
    Specification
    glioma cell, placenta, MDCK, HEK, HL-1
    Gel Running Conditions
    Reduced Denaturing (10)
    Blocking step
    FCS as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Mrs. Nicole Basler

    Verified customer

    Submitted Aug 04 2011

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

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