Overview

  • Product name
    Anti-MRP2 antibody [M2 III-6]
    See all MRP2 primary antibodies
  • Description
    Mouse monoclonal [M2 III-6] to MRP2
  • Host species
    Mouse
  • Specificity
    This antibody detects MRP 2. It does not cross-react with human MDR 1, MRP 1, MRP 3 or MRP 5 gene products.
  • Tested applications
    Suitable for: Flow Cyt, IP, ICC, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Dog
  • Immunogen

    Bacterial fusion protein of human MRP 2 containing the carboxy-terminal region (residues 1339-1541).

  • Epitope
    This antibody reacts with an internal epitope of MRP 2.
  • General notes

    We received mixed feedback about rat species so we are removing this from tested species list for now. The immunogen is 84% homologous to human protein so theoretically it should cross react however considering the latest customer feedback we no longer guarantee this species.

Properties

Applications

Our Abpromise guarantee covers the use of ab3373 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
Flow Cyt Use at an assay dependent concentration. PubMed: 18929567

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration. PubMed: 16687572
ICC 1/20 - 1/50.

Acetone fixed cytospin preparations.

IHC-P 1/200. See Abreview. Abreview indicates that several methods of antigen retreival are suitable for this antibody, but heat mediated Ag retrieval in citrate buffer gave optimal results.
IHC-Fr 1/20.

Acetone fixation of frozen sample is recommended.

WB 1/20 - 1/50. Predicted molecular weight: 174 kDa.

See Scheffer GL et al. 2000.

The samples MUST NOT be boiled before running the gel as multi-pass transmembrane proteins, like MRP2, aggregate severely at high temperatures, which increases as a direct function of temperature. We recommend heating the samples at 60-70C for 15-20 minutes or at 36C for 20 minutes as mentioned in PLoS Pathog 9:e1003244 (2013).
The protein is glycosylated at amino acid 7, 12 and 1011 so higher band size is expected.

Target

  • Function
    Mediates hepatobiliary excretion of numerous organic anions. May function as a cellular cisplatin transporter.
  • Tissue specificity
    Found on the apical membrane of polarized cells in liver, kidney and intestine. The highest expression is found in liver.
  • Involvement in disease
    Defects in ABCC2 are the cause of Dubin-Johnson syndrome (DJS) [MIM:237500]. DJS is an autosomal recessive disorder characterized by conjugated hyperbilirubinemia, an increase in the urinary excretion of coproporphyrin isomer I, deposition of melanin-like pigment in hepatocytes, and prolonged retention of sulfobromophthalein, but otherwise normal liver function.
  • Sequence similarities
    Belongs to the ABC transporter superfamily. ABCC family. Conjugate transporter (TC 3.A.1.208) subfamily.
    Contains 2 ABC transmembrane type-1 domains.
    Contains 2 ABC transporter domains.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ABC30 antibody
    • abcC2 antibody
    • ATP binding cassette sub family C (CFTR/MRP) member 2 antibody
    • ATP binding cassette subfamily C member 2 antibody
    • ATP-binding cassette sub-family C member 2 antibody
    • Canalicular multidrug resistance protein antibody
    • Canalicular multispecific organic anion transporter 1 antibody
    • CMOAT antibody
    • CMOAT1 antibody
    • cMRP antibody
    • DJS antibody
    • KIAA1010 antibody
    • MRP 2 antibody
    • MRP2_HUMAN antibody
    • Multidrug resistance associated protein 2 antibody
    • Multidrug resistance-associated protein 2 antibody
    see all

Images

  • Immunohistochemical analysis of Human liver tissue, staining MRP2 with ab3373 at 1/50 dilution.

References

This product has been referenced in:
  • Freyer N  et al. Microscale 3D Liver Bioreactor for In Vitro Hepatotoxicity Testing under Perfusion Conditions. Bioengineering (Basel) 5:N/A (2018). IHC ; Human . Read more (PubMed: 29543727) »
  • Jang M  et al. Study of melatonin-mediated effects on various hepatic inflammatory responses stimulated by IL-6 in a new HepG2-on-a-chip platform. Biomed Microdevices 20:54 (2018). Read more (PubMed: 29946898) »
See all 41 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell line)
Gel Running Conditions
Reduced Denaturing (8)
Loading amount
25 µg
Specification
Cancer Cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jun 28 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
15 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Armen Petrosyan

Verified customer

Submitted Jan 24 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Nov 17 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (WIF-B cells, the hepatoma-derived hybrid cell line)
Gel Running Conditions
Reduced Denaturing (6% SDS-PAGE)
Loading amount
30 µg
Specification
WIF-B cells, the hepatoma-derived hybrid cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 09 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepG2 cells)
Permeabilization
Yes - 0.2% Triton X-100
Specification
HepG2 cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Oct 14 2016

Answer


I can confirm that anti MRP2 antibody ab3373 does not cross react with other MRPs.

Read More

Question

Reviewing this case, I would like to offer some suggestions to help optimise the results from these antibodies. I would also appreciate if you can confirm some further details:
1. Please confirm the order numbers and dates of purchase.
Order reference number: #####

Dates: 06/08/2012
2. Could you confirm which lysis buffer was used? I can recommend to try RIPA buffer if not already tried. This should provide a suitable protein preparation.
The lysis buffer was a fixed kit, we bought from ThermoFischer to isolate membrane proteins and a buffer based on EDTA.
3. We recommend samples should be reduced and denatured and run on a reducing gel in WB unless stated otherwise on the datasheet. This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody.
The samples were reduced and diluted with the Laemmli buffer. Up to now I never used a reducing gel for my other ABC transporters and it always worked.
4. I can recommend to consider trying BSA rather than milk to block. Changing blocking agent can sometimes help to improve results.
This I will try
5. Could you confirm if the transfer to the membrane and quality of the samples been assessed using a loading control?
A coloured marker was used, showing a positive transfer onto the blot. A loading control was not used.
6. Are the current vials of secondary antibody and detection kit working well with other primary antibodies?
The detection kit works with other primary antibodies, the second antibody is new and didn't worked up to now. Can you maybe suggests an other working second antibody for these antibodies, I need with FITC labelling for IHC too.

Read More
Answer

Thank you for your reply and for kindly providing the requested details.

I look forward to hearing from you with the results of the next experiment. i can recommend it would be beneficial to consider a reducing gel. This will ensure the protein is in the correct conformation for detection by the antibody. Although the other antibodies work well using a non reducing gel, individual antibodies detect individual epitopes and so will sometimes require individual optimization.

I hope this will be helpful. Please do not hesitate to let me know how you get on with the reducing gel. I look forward to hearing from you.

Read More

Question

Thank you for your time and cooperation. I look forward to receiving the completed questionnaire.
Order Details
Antibody code:
MRP4: ab15602, GR79982-1
MRP5: ab 3377, GR43649-3
MRP2: ab 3373, GR19771-1
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
No signal in IHC for MRP4 and MRP5
Lot number
MRP4: ab15602, GR79982-1
MRP5: ab 3377, GR43649-3
MRP2: ab 3373, GR19771-1
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc)
-20oC except MRP5, fridge
Description of the problem (high background, wrong band size, more bands, no band etc.)
No band

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Cell extract

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Lyse Buffer with EDTA + Protease inhibitor cocktail

Amount of protein loaded
around 20 ug
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Non-reducing gel, 7.5% gel, running for 90 minutes, 150V, 80mA and 25W
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Buffer = without Methanol, with glycin and SDS for 2.5h with 320mA for two gels, blocking agent is a 2% Milkpowder in PBS+Tween20
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
MRP4: 1/50 dilution in milkpowder solution
MRP5: 1/50 dilution in milkpowder solution
MRP2: 1/100 dilution in milkpowder solution
Incubation in fridge overnight, washstep 3 times for 10 minutes with PBS+Tween20
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary antibody is from calbiochem, againt the whole IgG fraction. Produced in mouse or rat
MRP4: 1/100 dilution in milkpowder solution
MRP5: 1/50 dilution in milkpowder solution
MRP2: 1/100 dilution in milkpowder solution
Incubation 1Hour at roomtemperature, washstep 3 times for 10 minutes with PBS+Tween20
Detection method (ECL, ECLPlus etc.)
Peroxidase, chemilumiscence
Positive and negative controls used (please specify)
My positive control is the caco2 cell line, in which no detection occurs, no negative controls were used
Optimization attempts (problem solving)
How many times have you tried the Western?
Two times
Have you run a "No Primary" control?
No
Do you obtain the same results every time?
Yes
e.g. are the background bands always in the same place?
There are no background bands
What steps have you altered?
Non
Additional Notes:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.

Read More
Answer

Thank you for taking the time to complete our questionnaire. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from these antibodies. I would also appreciate if you can confirm some further details:

1. Please confirm the order numbers and dates of purchase.

2. Could you confirm which lysis buffer was used? I can recommend to try RIPA buffer if not already tried. This should provide a suitable protein preparation.

3. We recommend samples should be reduced and denatured and run on a reducing gel in WB unless stated otherwise on the datasheet. This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody.

4. I can recommend to consider trying BSA rather than milk to block. Changing blocking agent can sometimes help to improve results.

5. Could you confirm if the transfer to the membrane and quality of the samples been assessed using a loading control?

6. Are the current vials of secondary antibody and detection kit working well with other primary antibodies?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these antibodies.

I am sorry to confirm that because we carry over 90,000 products, it is regrettably not feasible for us to stock small sample sizes of our products to send to customers. If you are concerned about the secondary antibody, I can recommend to consider contacting the supplier of the secondary. I notice that MRP4 antibody [M4I-10] (ab15602) is a rat IgG2a, do you know that the secondary antibody will detect this isotype?

ab3373 Anti-MRP2 antibody [M2 III-6] should be working in WB, it has been tested successfully. Particularly if it has worked in the IHC experiments which shows the antibody is active, it is unusual this is not working in WB. Again, this is a mouse IgG2a isotype. Will the secondary antibody used in WB detect this, was this the same secondary used in IHC-P?

I would like to reassure you that all these antibodies are tested and covered by our 6 month guarantee for WB and the species that are listed on the individual datasheets. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

Thank you for your time and cooperation. I look forward to receiving the completed questionnaire.


Order Details
Antibody code:
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Yes No
Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?
What steps have you altered?
Additional Notes:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.

Read More

Answer



Any products that require an MSDS form will have a link on the product's online datasheet to the data. It will be a PDF document link labeled 'SDS'. Please see attached image for example of what to look for on the datasheet. If no 'SDS' link is available than the product does not contain any components that require an SDS.

Read More

1-10 of 17 Abreviews or Q&A

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

Sign up