MEK1 antibody (AA 2-150)

Catalog No. ABIN686482

Product Details anti-MEK1 Antibody

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

Binding Specificity: AA 2-150

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This MEK1 antibody is un-conjugated

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))

Cross-Reactivity: Human

Predicted Reactivity: Mouse,Rat,Dog,Pig,Chicken,Rabbit

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human MEK1

Isotype: IgG

Application Details

Application Notes: 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: 03/28/2014

Lot Number: 90915

Method validated: Immunohistochemistry

Positive Control: Human placenta

Negative Control: Human breast adipocytes

Notes: 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.

Full Methods

Primary Antibody:

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

Secondary Antibody:

• 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

Experimental 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.

Handling

Format: Liquid

Concentration: 1 μg/μL

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

Preservative: ProClin

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

Storage: 4 °C,-20 °C

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

Expiry Date: 12 months

Target Details for MEK1

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

Alternative Name: MEK1 (MAP2K1 Products)

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

Background:

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.

Gene ID: 5604

UniProt: Q02750

Pathways: MAPK Signaling, RTK Signaling, 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