Invitrogen

MEK1 Recombinant Polyclonal Antibody (22HCLC)

Advanced Verification

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

1 Published Figure

1 Reference

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Protocols

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MEK1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of MEK1 was done on 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with MEK1 Recombinant Rabbit Polyclonal Antibody (Product # 710446) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization of MEK1. Panel e shows no primary antibody control. The images were captured at 20X magnification.

MEK1 Antibody in Western Blot (WB) — Western blot analysis of MEK1 was performed by loading 20 µg of MCF7 (lane1) and A431 (lane2) cell lysates using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. MEK1 was detected at ~45 kDa using MEK1 Recombinant Rabbit Polyclonal Antibody (Product # 710446) at 1 µg-3 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

MEK1 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of MEK1 was done on NRK cells. Cells were fixed, permeabilized and stained with a MEK1/MAP2K1 Recombinant Rabbit Polyclonal Antibody (Product # 710446, blue histogram), or with a rabbit isotype control (green histogram) at 2 µg/million cells. After incubation for 1 hour on ice, the cells were labeled with DyLight 680 Goat anti-Rabbit Secondary Antibody (Product # 35568) at a dilution of 1:50 for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample. The black histogram represents unstained control cells and the red histogram represents no-primary-antibody control.

MEK1 Antibody — Evaluation of the antiviral hits CDK6, MAP2K1, and MAP2K3. 293/ACE2 cells were transfected with four individual siRNAs targeting, MAP2K1 (A and B), or MAP2K3 (C and D) or three CDK6-specific siRNAs (E and F). A nontargeting scrambled siRNA was used as a control. At 48 h p.t. cells were infected with SARS-CoV-GFP at an MOI of 10 (A, C, and E) and fixed 24 h later, and GFP fluorescence (black bars) was quantified and normalized to the value measured in infected, scrambled siRNA-transfected cells (100%). The effect of siRNA transfection on cell viability was analyzed in parallel (gray bars), and values were normalized to those of scrambled siRNA-transfected control cells (100%). Each experiment was repeated at least three times (averages ± standard deviations [SD]). In parallel, siRNA-transfected cells were infected with wt SARS-CoV (MOI, 5), and at 8 h p.i., SARS-CoV N expression was monitored by Western blotting (B, D, and F). TfR was used as a loading control. Knockdown levels of the host proteins were analyzed by Western blotting (B, D, and F). The amount of SARS-CoV N protein and remaining quantity of host protein compared to that of scrambled siRNA-transfected cells (100%) is shown below each lane. All experiments were repeated at least twice. {KD}

Product Details

710446

Applications	Tested Dilution	Publications
Western Blot (WB)	1-3 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	2-4 µg	-
Flow Cytometry (Flow)	2-4 µg/1x10^6 cells	-

Product Specifications
Species Reactivity	Human
Published species	Human
Host/Isotype	Rabbit / IgG
Expression System	EXPI293
Class	Recombinant Polyclonal
Type	Antibody
Clone	22HCLC
Immunogen	Peptide corresponding to amino acids 31–50 of human MEK1 View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS
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_2532717

Product Specific Information

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.

Product References

A Kinome-Wide Small Interfering RNA Screen Identifies Proviral and Antiviral Host Factors in Severe Acute Respiratory Syndrome Coronavirus Replication, Including Double-Stranded RNA-Activated Protein Kinase and Early Secretory Pathway Proteins.

Journal of virology

de Wilde AH,Wannee KF,Scholte FE,Goeman JJ,Ten Dijke P,Snijder EJ,Kikkert M,van Hemert MJ

710446 was used in western blot to identify proviral and antiviral host factors in SARS viral replication

Sat Aug 01 00:00:00 EDT 2015

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

Function(s)

MAPK cascade activation of MAPK activity movement of cell or subcellular component chemotaxis cell cycle arrest signal transduction heart development negative regulation of cell proliferation positive regulation of gene expression negative regulation of gene expression peptidyl-tyrosine phosphorylation cerebellar cortex formation neuron differentiation keratinocyte differentiation thyroid gland development regulation of stress-activated MAPK cascade proteolysis in other organism positive regulation of transcription, DNA-templated thymus development regulation of axon regeneration cell motility positive regulation of axonogenesis Bergmann glial cell differentiation face development trachea formation epithelial cell proliferation involved in lung morphogenesis placenta blood vessel development labyrinthine layer development ERK1 and ERK2 cascade positive regulation of ERK1 and ERK2 cascade positive regulation of protein serine/threonine kinase activity regulation of Golgi inheritance cellular senescence mitophagy in response to mitochondrial depolarization positive regulation of production of miRNAs involved in gene silencing by miRNA regulation of early endosome to late endosome transport

Process(es)

protein kinase activity protein serine/threonine kinase activity receptor signaling protein serine/threonine kinase activity MAP kinase kinase activity protein serine/threonine/tyrosine kinase activity protein tyrosine kinase activity receptor signaling protein tyrosine phosphatase activity protein binding ATP binding protein C-terminus binding protein kinase binding protein serine/threonine kinase activator activity protein N-terminus binding

MEK1 Recombinant Polyclonal Antibody (22HCLC)

Product Details

710446

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Expression System

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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