Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸

Order now and get it on Tuesday March 09, 2021

Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16667] to MEK1 + MEK2
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-MEK1 + MEK2 antibody [EPR16667]
See all MEK1 + MEK2 primary antibodies
Description

Rabbit monoclonal [EPR16667] to MEK1 + MEK2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Mouse
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Jurkat, Daudi, HeLa, 293T, A549 and A431 whole cell lysates; Human fetal brain, heart, kidney and spleen lysates; Mouse and Rat brain and heart lysates. IHC-P: Human renal medulla, Mouse lung and Rat lung tissues. ICC/IF: NIH/3T3 cells. IP: HeLa whole cell extract.
General notes
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16667
Isotype

IgG
Research areas

Western blot - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

All lanes : Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876) at 1/20000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : MAP2K2 knockout HEK293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : Daudi cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 43, 44 kDa
Observed band size: 45 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab178876 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab178876 was shown to react with MEK2 in wild-type HEK-293T cells in western blot with loss of signal observed in MAP2K2 knockout sample. Wild-type HEK-293T and MAP2K2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab178876 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

ab178876 staining MEK1 + MEK2 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Immunocytochemistry/ Immunofluorescence - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling MEK1 + MEK2 with ab178876 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/200 dilution (green). Cytoplasmic staining on NIH/3T3 cell line is observed. The nuclear counter stain 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. ab178876 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 1/200 dilution.
Western blot - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

All lanes : Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876) at 1/20000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates
Lane 3 : A549 (Human lung carcinoma) whole cell lysates
Lane 4 : A431 (Human epidermoid carcinoma) whole cell lysates

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 43, 44 kDa
Observed band size: 44 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

All lanes : Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876) at 1/50000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lane 4 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 43, 44 kDa
Observed band size: 44 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

All lanes : Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876) at 1/50000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : Mouse heart lysate
Lane 4 : Rat heart lysate

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: 43, 44 kDa
Observed band size: 44 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

Immunohistochemical analysis of paraffin-embedded Human renal medulla tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Renal tubule epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

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-MEK1 + MEK2 antibody [EPR16667] (ab178876)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Alveolar epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

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-MEK1 + MEK2 antibody [EPR16667] (ab178876)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Alveolar epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

MEK1 + MEK2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab178876 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab178876 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 and concentration: 5% NFDM/TBST.
Anti-MEK1 + MEK2 antibody [EPR16667] (ab178876)

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