Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free
See all MEK3 + MEK6 primary antibodies
Description

Rabbit monoclonal [EPR17345] to MEK3 + MEK6 - BSA and Azide free
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-P, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human, Recombinant fragment
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
General notes
Ab250507 is the carrier-free version of ab181555. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab250507 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17345
Isotype

IgG
Research areas

Western blot - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/50000 dilution

Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 37, 39 kDa
Observed band size: 39,37,34 kDa
why is the actual band size different from the predicted?

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.
Immunocytochemistry - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling MEK3 + MEK6 with ab181555 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:-
1. ab181555 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
Western blot - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/10000 dilution + Human MEK6 recombinant protein fragment at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 37, 39 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Recombinant protein covers 139aa-334aa with a His tag (22kDa).
Western blot - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/5000 dilution

Lane 1 : Rat liver lysates
Lane 2 : Mouse liver lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 37, 39 kDa
Observed band size: 39,37,34 kDa why is the actual band size different from the predicted?

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/2000 dilution

Lane 1 : Mouse heart lysates
Lane 2 : Mouse kidney lysates
Lane 3 : Rat heart lysates
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 37, 39 kDa
Observed band size: 39,37,34 kDa why is the actual band size different from the predicted?

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Human skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Mouse skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Rat skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary ab. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab181555 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)

This data was developed using ab181555, the same antibody clone in a different buffer formulation.

MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab33866 (Rabbit monoclonal [EP557Y] to MEK6) at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.

Blocking and dilution buffer: 5% NFDM/TBST.
Anti-MEK3 + MEK6 antibody [EPR17345] - BSA and Azide free (ab250507)