Invitrogen

Phospho-MEK1 (Thr386) Recombinant Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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Phospho-MEK1 (Thr386) Antibody in Immunocytochemistry (ICC/IF) — For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous Mek1 pT386 using Anti- Mek1 pT386 Recombinant Rabbit Polyclonal Antibody (Product # 711612, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Mek1 pT386 (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating localization of Mek1 pT386 in dividing cells. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Phospho-MEK1 (Thr386) Antibody in Western Blot (WB) — Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1) and HEK-293 treated with EGF (100 ng/mL for 30 mins) (Lane 2). The blots were probed with Anti-MEK1 (pT386) Recombinant Rabbit Polyclonal Antibody (Product # 711612, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 43 kDa band corresponding to MEK1 (pT386) was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Phospho-MEK1 (Thr386) Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot of MEK1 [pT386] using Anti-MEK1 [pT386] Recombinant Rabbit Polyclonal Antibody (Product # 711612), shows expression of MEK1 [pT386] in HEK-293 upon treatment with EGF. {TM}

Product Details

711612

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	-
Immunocytochemistry (ICC/IF)	2 µg/mL	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Polyclonal
Type	Antibody
Immunogen	Peptide corresponding to Human MAP2K1 (aa Chemically synthesized phosphopeptide derived from a region of human MEK1 that contains threonine 386. The sequence is conserved in many species including mouse, rat, chimp, hamster, and rabbit. (Antigen similar to 44-462G ))
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.4
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2632983

Product Specific Information

This antibody is predicted to react with Monkey, Bovine and Pig.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

MEK1/mitogen activated protein kinase/ERK kinase 1/microtubule-associated protein 2 kinase is a dual specificity kinase and catalyzes both a threonine and a tyrosine residue on MAP kinases ERK1 and ERK2. Mek1 is activated by phosphorylation of serine 218 and 222 residues by Raf1. It is known to be involved in the signaling during stress activate response, apoptosis and proliferative induction by cytokines. Mutations in the gene can lead to cardiofaciocutaneous syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Dual specificity mitogen-activated protein kinase kinase 1; EC 2.7.12.2; ERK activator kinase 1; kinase MEK1; MAP kinase kinase 1; MAPK/ERK kinase 1; MAPKK 1; MEK 1; MKK (Thr292); MKK (Thr386); MKK2; MP2K1; PRKMK2; protein kinase, mitogen-activated, kinase 1 (MAP kinase kinase 1)

Gene Aliases: CFC3; MAP2K1; MAPKK1; MEK1; MKK1; PRKMK1

UniProt ID: (Human) Q02750

Entrez Gene ID: (Human) 5604

Molecular Function: non-receptor serine/threonine protein kinase