Phospho-MEK1/2 (S218 + S222) | HUABIO ET1609-50 product information

product summary

company name :

HUABIO

product type :

antibody

product name :

Phospho-MEK1/2 (S218 + S222)

catalog :

ET1609-50

quantity :

100μl

price :

385.00 USD

clonality :

monoclonal

host :

domestic rabbit

conjugate :

nonconjugated

clone name :

ST0490

reactivity :

human, mouse, rat

application :

western blot, immunoprecipitation, immunohistochemistry - paraffin section

more info or order :

HUABIO product webpage

image

image 1 :

Western blot analysis of Phospho-MEK1/2 (S218 + S222) on different lysates with Rabbit anti-Phospho-MEK1/2 (S218 + S222) antibody (ET1609-50) at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate Lane 3: C6 cell lysate Lane 4: C6 treated with 200nM PMA for 30 minutes cell lysate Lane 5: NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lane 6: C6 treated with 200nM PMA for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 43 kDa Observed band size: 43 kDa Exposure time: 24 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 (ET1609-50) at 1/1,000 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.

image 2 :

Western blot analysis of Phospho-MEK1/2 (S218 + S222) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1609-50, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hela cell lysate Lane 2: A431 cell lysate Lane 2: 293T cell lysate

image 3 :

Western blot analysis of Phospho-MEK1/2 (S218 + S222) on Hela cell lysates. Lane 1: Hela cells, whole cell lysate, 10ug/lane Lane 2/3: Hela cells treated with 200 nM PMA for 20 minutes, whole cell lysates, 10ug/lane Lane 4: Hela cells treated with 200 nM PMA for 20 minutes, then treated with 2.8ug/ul lambda-PP for 30 minutes, whole cell lysates, 10ug/lane All lanes : Anti-Phospho-MEK1/2 (S218 + S222) antibody (ET1609-50) at 1/500 dilution. Anti-MEK1 antibody (ET1603-20) at 1/500 dilution. Anti-GAPDH antibody (ET1601-4) at 1/10,000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (HA1001) at 1/200,000 dilution. Predicted band size:43 kDa Observed band size:43 kDa Blocking and diluting buffer: 5% BSA. Exposure time: Lane1/2 5 minutes Lane3/4 1 minutes 32 seconds

product information

SKU :

ET1609-50

Target name :

Phospho-MEK1/2 (S218 + S222)

Species reactivity :

Human,Mouse,Rat

Applications :

WB,IF-Cell,IF-Tissue,IHC-P,IP

Conjugate :

Non-conjugated

Immunogen :

Synthetic phospho-peptide corresponding to residues surrounding Ser218 and 222 of Human MEK1 aa 200-249 / 393.

Uniprot id :

Q02750>SwissProt: Q02750 Human;SwissProt: P31938 Mouse;SwissProt: Q01986 Rat

Host :

Rabbit

Clone number :

ST0490

Isotype :

IgG

Size :

100μl

List Price :

385.00 USD

Storage Buffer :

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Form :

Liquid

Storage Instruction :

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Purity :

Protein A affinity purified.

Product type :

Recombinant Rabbit monoclonal Antibody

Positive control :

NIH/3T3 treated with 200nM PMA for 30 minutes cell lysate, C6 treated with 200nM PMA for 30 minutes cell lysate, Hela cell lysate, A431 cell lysate, 293T cell lysate, NIH/3T3 cells treated with 200nM PMA for 30 minutes, Hela, HepG2, human tonsil tissue, human liver carcinoma tissue, human spleen tissue, human breast carcinoma tissue.

Molecular wt :

Predicted band size: 43 kDa

Subcellular location :

Cytoplasm, Cytoskeleton, Membrane, Nucleus.

Concentration :

1 mg/mL.

Recommended dilutions :

WB: 1:500-1:2,000 ;IF-Cell: 1:50-1:200 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:50-1:200 ;IP: Use at an assay dependent concentration.

Advanced Validation :

Cell treatment (CT)

Pic img4 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_4.jpg

Pic legend4 :

Immunocytochemistry analysis of NIH/3T3 cells treated with 200nM PMA for 30 minutes labeling Phospho-MEK1/2 (S218 + S222) with Rabbit anti-Phospho-MEK1/2 (S218 + S222) antibody (ET1609-50) 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-Phospho-MEK1/2 (S218 + S222) antibody (ET1609-50) 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.

Pic img5 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_5.jpg

Pic legend5 :

ICC staining of Phospho-MEK1/2 (S218 + S222) in Hela 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 (ET1609-50, 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).

Pic img6 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_6.jpg

Pic legend6 :

ICC staining of Phospho-MEK1/2 (S218 + S222) in HepG2 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 (ET1609-50, 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).

Pic img7 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_7.jpg

Pic legend7 :

Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-MEK1/2 (S218 + S222) 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 (ET1609-50, 1/50) 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.

Pic img8 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_8.jpg

Pic legend8 :

Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Phospho-MEK1/2 (S218 + S222) 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 (ET1609-50, 1/50) 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.

Pic img9 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_9.jpg

Pic legend9 :

Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Phospho-MEK1/2 (S218 + S222) 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 (ET1609-50, 1/50) 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.

Pic img10 :

https://storage.huabio.cn/huabio/productImg/ET1609-50_10.jpg

Pic legend10 :

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-MEK1/2 (S218 + S222) 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 (ET1609-50, 1/50) 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.

more info or order :

HUABIO product webpage