Overview

  • Product name

    Anti-SLC16A3/MCT 4 antibody
    See all SLC16A3/MCT 4 primary antibodies
  • Description

    Rabbit polyclonal to SLC16A3/MCT 4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ELISAmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human SLC16A3/MCT 4 (internal sequence).

  • Positive control

    • Extracts from LOVO cells
  • General notes

    Protein previously labeled as SLC16A3.

Properties

Applications

Our Abpromise guarantee covers the use of ab74109 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 1/500 - 1/1000. Predicted molecular weight: 49 kDa.
ELISA 1/40000.

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

    Highly expressed in skeletal muscle.
  • 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

  • Form

    Lactic acid and pyruvate transport across plasma membranes is catalyzed by members of the proton linked monocarboxylate transporter (MCT) family, which has been designated solute carrier family 16. Each MCT appears to have slightly different substrate and inhibitor specificities and transport kinetics, which are related to the metabolic requirements of the tissues in which it is found. The MCTs, which include MCT1 and MCT2, are characterized by 12 predicted transmembrane domains.
  • Alternative names

    • MCT 4 antibody
    • MCT3 antibody
    • MCT4 antibody
    • Monocarboxylate transporter 3 antibody
    • Monocarboxylate transporter 4 antibody
    • MOT4_HUMAN antibody
    • SLC16A3 antibody
    • Solute carrier family 16 member 3 antibody
    see all

Images

  • All lanes : Anti-SLC16A3/MCT 4 antibody (ab74109) at 1/500 dilution

    Lane 1 : extracts from LOVO cells
    Lane 2 : extracts from LOVO cells with immunizing peptide at 5 µg

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 49 kDa
    Observed band size: 49 kDa

References

This product has been referenced in:

  • Sun K  et al. Oxidized ATM-mediated glycolysis enhancement in breast cancer-associated fibroblasts contributes to tumor invasion through lactate as metabolic coupling. EBioMedicine 41:370-383 (2019). Read more (PubMed: 30799198) »
  • Zawislak A  et al. Neuron-derived transthyretin modulates astrocytic glycolysis in hormone-independent manner. Oncotarget 8:106625-106638 (2017). Read more (PubMed: 29290976) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Chicken Tissue lysate - whole (Breast)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
Breast
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 26 2017

Application
Western blot
Sample
Pig Tissue lysate - whole (Skeletal Muscle)
Gel Running Conditions
Reduced Denaturing (10% 37.5:1)
Loading amount
30 µg
Specification
Skeletal Muscle
Blocking step
Thermo Starting Block 37542 as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 10 2013

Question

WB Questionnaire

It will take approximately 5 minutes to complete the questionnaire.

Please fill-in all fields so that we can better assist you

1) Abcam product code

MCT4 ab74109

2) Abcam order reference number or product batch number:

953528

3) Description of the problem

For the MCT4 I got one specific and strong band in my samples, however, it was not the size you predict (46kDa). The band on my blot is between 52 and 76kDa.

4) Sample preparation:

Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysates

Lysis buffer Laemmli

Protease inhibitors: Protease inhibitor cocktail (Sigma, P8340)

Phosphatase inhibitors

Reducing agent: SDS

Boiling for ≥5 min? yes/no: yes

Protein loaded ug/lane or cells/lane 15 µg or 20µg

Positive control: MDA-MB 231 cells

Negative control: none

5) Percentage of gel 4-12% gradient gel, 10%, 12%

Type of membrane nitrocellulose

Protein transfer verified: yes, ponceau staining

Blocking agent and concentration 5% milk in TBS-t

Blocking time: 30 min

Blocking temperature: room temperature

6) Was a fresh membrane used or had it been probed and stripped with a different antibody?

Membrane was stripped this time, but formerly in another experiment a fresh membrane was used

7) Primary antibody (If more than one was used, describe in “additional notes”) :

- 1st antibody: LDHA (ab92903) 1 in 1000, membrane stripped

- 1st antibody: LDHB, 1 in 1000, membrane stripped

- 2nd antibody MCT4 (ab74109), 1 in 500. Antibody was formerly used 1 in 1000 but with no results

Detailed description see ppt of WB attached

Concentration or dilution: 1 in 1000, 1 in 500

Diluent buffer: 5% mil in TBS-T

Incubation time over night, 16 hours

Incubation temperature: 4˚C

8) Secondary antibody: LICOR 800CW or LICOR 680

Species:

- MCT4 → rabbit

Reacts against:

Concentration or dilution: 1 in 4000

Diluent buffer: 5% mlik in TBST

Incubation time: 45 min

Incubation temperature: room temp

Fluorochrome or enzyme conjugate: LICOR 800CW, LICOR 680

9) Washing after primary and secondary antibodies:

Buffer: TBST

Number of washes 3x 5 min each

10)Detection method: LICOR fluorescent detection

11) How many times have you run this staining? 6 times

Do you obtain the same results every time? No, the first time no bands were visible

What steps have you altered to try and optimize the use of this antibody?

The first time antibody was used 1 in 1000, 2nd time 1 in 500

I also tried HRP coupled secondary antibodies instead of LICOR, but obtained no bands.

Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Read More
Answer

Thank you for providing that information. It allows me to more fully understand the results that you have been observing.

There are a few further things I would like to clarify:

You mentioned that the anti-MCT1 has also been proving difficult to get the expected results? Could you describe this more fully? Possibly filling out the questionnaire in again as you have done for ab74109 and providing the data observed?

Did the initial incubation with the anti-LDHA and LDHB yield the expected band sizes?

It seems that the unsexpected band size is only observed when the membrane has been stripped and re-probed. It may be that the stripping process is not 100% efficient and the secondary antibody is picking up the first antibody used, not ab74109. It does however, appear that without stripping being performed there is no band, which would not be the expected results with the protocol you have been using and the samples. It may therefore be that the vial of this antibody which you have recieved is faulty. I am very sorry for this and for the inconvinience it has caused you. I am able to offer you a free of charge repacement with a different vial. Or if you would prefer I can arrange for a refund or credit note.

I look forward to hearing how you would like to proceed.

Read More

Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties withtwo of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I have had a look into the information we have regarding ab74109 and ab90582 but cannot suggest anything immediately that may be the cause of the problem. In order to more fully understand what may be contributing to the results observed I haveattacheda questionnaire so thatI can gather further information regarding the samples tested and the protocol used. OnceI have receive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.


If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More

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