MEK1/2 Recombinant Rabbit Monoclonal Antibody [SR13-07] - BSA and Azide free
Catalog# HA750042
MEK1/2 Recombinant Rabbit Monoclonal Antibody [SR13-07] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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ET1602-3
含抗保成分
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Zebrafish
Predicted species support after-sales service
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unconjugated
Overview
Product Name
MEK1/2 Recombinant Rabbit Monoclonal Antibody [SR13-07] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human MEK1 aa 1-393 / 393.
Species Reactivity
Human, Mouse, Rat (Predicted: Zebrafish)
Predicted species support after-sales service
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 44 kDa
Positive Control
HeLa cell lysate, HepG2 cell lysate, A431 cell lysate, NIH/3T3 cell lysate, mouse brain tissue lysate, PC-12 cell lysate, A549, NIH/3T3, human kidney tissue, mouse lung tissue, mouse kidney tissue, mouse hippocampus tissue, mouse brain tissue.
Conjugation
unconjugated
Clone Number
SR13-07
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:50
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IF-Tissue
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1:50
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IHC-P
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1:200
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IP
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1-2μg/sample
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FC
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1:1,000
Target
Function
A family of protein kinases located upstream of the MAP kinases and responsible for their activation has been identified. The prototype member of this family, designated MAP kinase kinase, or MEK-1, specifically phosphorylates the MAP kinase regulatory threonine and tyrosine residues present in the Thr-Glu-Tyr motif of ERK. A second MEK family member, MEK-2, resembles MEK-1 in its substrate specificity. MEK-3 (or MKK-3) functions to activate p38 MAP kinase, and MEK-4 (also called SEK1 or MKK-4) activates both p38 and JNK MAP kinases. MEK-5 appears to specifically phosphorylate ERK5, whereas MEK-6 phosphorylates p38 and p38b. MEK-7 (or MKK-7) phosphorylates and activates the JNK signal transduction pathway.
Background References
1. Jeong, Y. et al. 2014. Histone deacetylase isoforms regulate innate immune responses by deacetylating mitogen-activated protein kinase phosphatase-1. Journal of leukocyte biology. 95: 651-9.
2. Zhang, M. et al. 2013. CD133 affects the invasive ability of HCT116 cells by regulating TIMP-2. Am. J. Pathol. 182: 565-576.
Sequence Similarity
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.
Post-translational Modification
MAPKK is itself dependent on Ser/Thr phosphorylation for activity catalyzed by MAP kinase kinase kinases (RAF or MEKK1). Phosphorylated by MAP2K1/MEK1 (By similarity).; Acetylation of Ser-222 and Ser-226 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.
Subcellular Location
Cytoplasm, Nucleus, Membrane, Mitochondrion
UNIPROT
Synonyms
AA589381 antibody
CFC3 antibody
Dual specificity mitogen-activated protein kinase kinase 1 antibody
Dual specificity mitogen-activated protein kinase kinase 2 antibody
EC 2.7.12.2 antibody
ERK activator kinase 1 antibody
ERK activator kinase 2 antibody
FLJ26075 antibody
MAP kinase kinase 1 antibody
MAP kinase kinase 2 antibody
ExpandImages
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Western blot analysis of MEK1/2 on different lysates with Rabbit anti-MEK1/2 antibody (HA750042) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HepG2 cell lysate (15 µg/Lane)
Lane 3: A431 cell lysate (15 µg/Lane)
Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: PC-12 cell lysate (15 µg/Lane)
Predicted band size: 44 kDa
Observed band size: 44 kDa
Exposure time: 1 minute 21 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750042) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of MEK1/2 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750042, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of MEK1/2 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750042, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-MEK1/2 antibody (HA750042) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-MEK1/2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-MEK1/2 antibody (HA750042) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-MEK1/2 antibody (HA750042) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-MEK1/2 antibody (HA750042) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of PC-12 cells labeling MEK1/2 with Rabbit anti-MEK1/2 antibody (HA750042) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MEK1/2 antibody (HA750042) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MEK1/2 antibody (HA750042) at 1/10,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750042) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of A549 cells labeling MEK1/2.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750042, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
MEK1/2 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA750042 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750042 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA750042 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA750042 in HeLa cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Products with the same target and pathway
MEK1/2 Recombinant Rabbit Monoclonal Antibody [SR13-07]
Application: WB,IF-Cell,IF-Tissue,IHC-P,IP,FC
Reactivity: Human,Mouse,Rat,Zebrafish
Conjugate: unconjugated
