WB result of MEK1/2 Recombinant Rabbit mAb
Primary antibody: MEK1/2 Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: A431 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 45 kDa
MEK1/2 Recombinant Rabbit mAb (S-834-101)
MEK1/2 Recombinant Rabbit mAb (S-834-101)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | MEK1/2 |
| Synonyms | Dual specificity mitogen-activated protein kinase kinase 1; MAP kinase kinase 1; MAPKK 1; MKK1; ERK activator kinase 1; MAPK/ERK kinase 1 |
| Immunogen | Recombinant Protein |
| Location | Nucleus, Cytoskeleton, Cytoplasm, Membrane |
| Accession | Q02750 |
| Clone Number | S-834-101 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM, IP |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:5000 | |
| IP | 1:50 | |
| IHC-P | 1:250 | |
| ICC | 1:500 | |
| ICFCM | 1:500 |
Background
MEK1/2 (Mitogen-Activated Protein Kinase Kinase 1 and 2) are crucial components within the MAPK (Mitogen-Activated Protein Kinase) signaling pathway, belonging to the protein kinase family. They are activated by upstream kinases such as RAF, which then phosphorylate and activate downstream ERK (Extracellular Signal-Regulated Kinase) protein kinases, thereby promoting cellular processes like growth, differentiation, and proliferation. In many cancers, MEK1/2 activity is significantly elevated, enabling tumor cells to proliferate and grow unchecked. Therefore, inhibiting the MEK1/2 signaling pathway is considered a vital anticancer strategy.
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Western Blot
WB result of MEK1/2 Recombinant Rabbit mAb
Primary antibody: MEK1/2 Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 45 kDa
WB result of MEK1/2 Recombinant Rabbit mAb
Primary antibody: MEK1/2 Recombinant Rabbit mAb at 1/5000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 45 kDa
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) labelling MEK1/2 antibody at 1/500 dilution (0.1 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
IP
MEK1/2 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating MEK1/2 in 0.4 mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using MEK1/2 Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 20 µg (Input)
Lane 2: MEK1/2 Rabbit mAb IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 43 kDa
Observed MW: 45 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-MEK1/2 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human kidney. Anti-MEK1/2 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human liver. Anti-MEK1/2 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-MEK1/2 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-MEK1/2 antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-MEK1/2 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ICC shows positive staining in NIH/3T3 cells. Anti-MEK1/2 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
