MEK1 anticorps (AA 2-150)

N° du produit ABIN686482

Détail du produit anti-MEK1 anticorps

Antigène: MEK1 (MAP2K1) (Mitogen-Activated Protein Kinase Kinase 1 (MAP2K1))

Épitope: AA 2-150

Reactivité: Humain

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp MEK1 est non-conjugé

Application: ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

Réactivité croisée: Humain

Homologie: Mouse,Rat,Dog,Pig,Chicken,Rabbit

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic peptide derived from human MEK1

Isotype: IgG

Information d'application

Indications d'application: ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029646 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029646

Date: 28.03.2014

Numéro du lot: 90915

Application validée: Immunohistochemistry

Contrôle positif: Human placenta

Contrôle négative: Human breast adipocytes

Conclusion: Signal detected in positive control sample and not in negative control sample.

Validation Images

Figure 1: Human placenta tissue stained with anti-MAP2K1 (brown) and counterstained with hematoxylin.

Figure 2: Human breast adipose tissue stained with anti-MAP2K1 (brown) and counterstained with hematoxylin.

Figure 3: Human placenta tissue stained with isotype control antibody (brown) and counterstained with hematoxylin.

Figure 4: Human placenta tissue stained with secondary antibody only (brown) and counterstained with hematoxylin.

Protocole

Anticorps primaire:

• Antigen: Mitogen-Activated Protein Kinase Kinase 1 (MAP2K1)
• Catalog number: ABIN686482
• Lot number: 90915

Anticorps secondaire:

• Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
• Lot number: D07640BA

Full Protocol:

• Immunohistochemistry was performed on a Ventana NEXes automated platform; instrument manufacturer specific reagents are italicized.
• 1. Slides were preheated in convection oven at 60°C for 30 min
• 2. Deparaffinization procedure: - 3 changes of Xylene, 5 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of 95% Ethanol, 3 min each - Rinsed in distilled water, 3 changes
• 3. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 min. - Slides were allowed to cool (in citrate) for 30 min. - Slides were washed x 3 in Distilled water
• 4. NEXes instrument procedure, iView DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
• 5. Slides rinsed with *reaction buffer* x3
• 6. *iView Inhibitor (H2O2)* applied and incubated for 4 min
• 7. Slides rinsed with *reaction buffer*
• 8. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliter applied/slide) - Ventana Isotype control applied neat - Slides Incubated overnight at room temperature (~12 hours ~25°C)
• 9. Slides rinsed with *reaction buffer* x3
• 10. *iView Biotinylated IgG* applied and incubated for 8 min
• 11. Slides rinsed with *reaction buffer*
• 14. *iView Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
• 15. Slides rinsed with *reaction buffer*
• 16. *iView DAB/H2O2* applied and incubated for 8 min
• 17. Slides rinsed with *reaction buffer*
• 18. *iView Copper* applied and incubated for 4 min
• 19. Slides rinsed with *reaction buffer*
• 20. Slides washed in Dawn Detergent/tap water
• 21. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 sec - Slides washed in tap water, 1 min - Decolorized (10% Acetic Acid in 70% ethanol), 1 min - Slides washed in tap water, 1 min - Bluing (Austin Clear Ammonia), 1 min - Slides washed in tap water, 1 min
• 22. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of Xylene, 5 min each - Mounted with Permount
• 23. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software

Notes:

• Deviations from protocol/procedure supplied by manufacturer:
• Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
• Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
• Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
• Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
• Step 4: Italicized reagents and incubation time are fixed instrument parameters.
• Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
• Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
• Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.

Stockage

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Agent conservateur: ProClin

Précaution d'utilisation: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Stock: 4 °C,-20 °C

Stockage commentaire: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Date de péremption: 12 months

Détails sur MEK1

Antigène: MEK1 (MAP2K1) (Mitogen-Activated Protein Kinase Kinase 1 (MAP2K1))

Autre désignation: MEK1 (MAP2K1 Produits)

Synonymes: anticorps Mek1, anticorps CFC3, anticorps MAPKK1, anticorps MEK1, anticorps MKK1, anticorps PRKMK1, anticorps MEKK1, anticorps Prkmk1, anticorps mek-2, anticorps si:ch211-242m18.2, anticorps wu:fj56a12, anticorps wu:fj61b01, anticorps zgc:56557, anticorps ATMEK1, anticorps F20B18.180, anticorps F20B18_180, anticorps MAP kinase/ ERK kinase 1, anticorps MITOGEN ACTIVATED PROTEIN KINASE KINASE, anticorps MITOGEN ACTIVATED PROTEIN KINASE KINASE 1, anticorps NMAPKK, anticorps MEK, anticorps mitogen activated protein kinase kinase 1, anticorps mitogen-activated protein kinase kinase 1, anticorps mitogen-activated protein kinase kinase 1 L homeolog, anticorps MAP kinase/ ERK kinase 1, anticorps Map2k1, anticorps MAP2K1, anticorps map2k1.L, anticorps map2k1, anticorps MEK1

Sujet:

Synonyms: CFC3, MEK1, MKK1, MAPKK1, PRKMK1, Dual specificity mitogen-activated protein kinase kinase 1, MAP kinase kinase 1, MAPKK 1, ERK activator kinase 1, MAPK/ERK kinase 1, MEK 1, MAP2K1

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.

ID gène: 5604

UniProt: Q02750

Pathways: Signalisation MAPK, Signalisation RTK, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Toll-Like Receptors Cascades, Autophagy, Signaling of Hepatocyte Growth Factor Receptor, BCR Signaling