MEK1 Antikörper (AA 2-150)

Produktnummer ABIN686482

Produktdetails anti-MEK1 Antikörper

Highlights:

• High quality MEK1 Primärantikörper for the detection of MAP2K1.
• Reliable product with high standard validation data

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

Bindungsspezifität: AA 2-150

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser MEK1 Antikörper ist unkonjugiert

Applikation: 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))

Kreuzreaktivität: Human

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

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human MEK1

Isotyp: IgG

Produktspezifische Information:

What can the MEK1 Antibody ABIN686482 be used for?: This polyclonal MEK1 Antibody detects MEK1. The MEK1 Antibody has been validated for various applications and can be used for the detection of MEK1 and derivatives by ELISA, Immunohistochemistry (Paraffin-embedded Sections), Immunofluorescence (Cultured Cells), Immunofluorescence (Paraffin-embedded Sections), Flow Cytometry, Immunohistochemistry (Frozen Sections).

What validation data is available for this MEK1 Antibody?: The Primärantikörper has currently 3 product images that show its performance in a variety of applications. The product is currently available in 100 μL quantity. MEK1 Antibody for the detection of MEK1 and derivatives.

What is the function of MEK1?: The protein encoded by this gene is a member of the dual specificity protein kinase family, which acts as a mitogen-activated protein (MAP) kinase kinase. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act as an integration point for multiple biochemical signals. This protein kinase lies upstream of MAP kinases and stimulates the enzymatic activity of MAP kinases upon wide variety of extra- and intracellular signals. As an essential component of MAP kinase signal transduction pathway, this kinase is involved in many cellular processes such as proliferation, differentiation, transcription regulation and development. [provided by RefSeq, Jul 2008].

Anwendungsinformationen

Applikationshinweise: 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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #029646 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029646

Datum: 28.03.2014

Chargennummer: 90915

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: Human placenta

Negativkontrolle: Human breast adipocytes

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

Validierungsbilder

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.

Protokoll

Primärantikörper:

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

Sekundärantikörper:

• 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

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

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

Konservierungsmittel: ProClin

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

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Details zu MEK1

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

Andere Bezeichnung: MEK1 (MAP2K1 Produkte)

Hintergrund:

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.

Gen-ID: 5604

UniProt: Q02750

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