MEK1 (S298) Monoclonal Antibody

$25 ug total protein per lane of various lysates (see on figure) probed with phospho-MEK1 (Ser298) monoclonal antibody, unconjugated (bsm-52175R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with phospho-MEK1 (Ser298) monoclonal antibody, unconjugated (bsm-52175R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$A431 (H) treated with nocodazole (100ng\/ml) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MEK1 (Ser298) monoclonal antibody, unconjugated (bsm-52175R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

A431 (H) treated with nocodazole (100ng\/ml) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MEK1 (Ser298) monoclonal antibody, unconjugated (bsm-52175R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$Paraformaldehyde-fixed, paraffin embedded Mouse Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-MEK1 (Ser298) Monoclonal Antibody, Unconjugated (bsm-52175R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MEK1 (Ser298)) monoclonal Antibody, unconjugated (bsm-52175R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed HeLa (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MEK1 (Ser298)) monoclonal Antibody, unconjugated (bsm-52175R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-MEK1 (Ser298) antibody (bsm-52175R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The HeLa (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-MEK1 (Ser298) antibody (bsm-52175R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

Applications

WB
IF(ICC)

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-52175r
Product Name	MEK1 (S298) Monoclonal Antibody
Applications	WB IF(ICC)
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	A synthesized peptide derived from human MEK1 around the phosphorylation site of S298
Modification Site	S298
Clonality	Recombinant
Clone #	20B10
Isotype	IgG
Concentration	1mg/ml
Purification	purified by Protein A.
Storage Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	5604
Swiss Prot	Q02750
Synonyms	MAP2K2 \| MEK1 (phospho-S298); p-MEK1; phospho-MEK1; p-MAP2K2; MAP2K2 \| MEK1 (phospho-Ser298); MP2K1_HUMAN; MAP2K1; MAP kinase kinase 1; MAPKK 1; MKK1; ERK activator kinase 1; MAPK/ERK kinase 1 (MEK 1); 2.7.12.2; MEK1; PRKMK1; CFC3; MAPKK1; MEL;
Background	Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. RAF1 then further activates the dual-specificity protein kinases MAP2K1/MEK1 and MAP2K2/MEK2. Both MAP2K1/MEK1 and MAP2K2/MEK2 function specifically in the MAPK/ERK cascade, and catalyze the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, leading to their activation and further transduction of the signal within the MAPK/ERK cascade. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth, adhesion, survival and differentiation, predominantly through the regulation of transcription, metabolism and cytoskeletal rearrangements. One target of the MAPK/ERK cascade is peroxisome proliferator-activated receptor gamma (PPARG), a nuclear receptor that promotes differentiation and apoptosis. MAP2K1/MEK1 has been shown to export PPARG from the nucleus. The MAPK/ERK cascade is also involved in the regulation of endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC), as well as in the fragmentation of the Golgi apparatus during mitosis.
Application Dilution
WB	1:300-5000
IF(ICC)	1:50-200