MAP2K5 antibody (Mitogen-Activated Protein Kinase Kinase 5) (AA 380-430)

Details for Product anti-MAP2K5 Antibody No. ABIN754183, Supplier: Log in to see
Antigen
  • Mek5
  • HsT17454
  • MAPKK5
  • MEK5
  • PRKMK5
  • AI324775
  • AI428457
  • Mapkk5
  • Prkmk5
  • fb73f02
  • wu:fb73f02
  • zgc:172137
  • ARABIDOPSIS THALIANA MITOGEN-ACTIVATED PROTEIN KINASE KINASE 5
  • ATMAP2K_ALPHA
  • ATMEK5
  • ATMKK5
  • MAP KINASE KINASE 5
  • MAP kinase kinase 5
  • MAP2K_A
  • mitogen activated protein kinase kinase 5
  • mitogen-activated protein kinase kinase 5
  • Map2k5
  • MAP2K5
  • map2k5
  • MKK5
Alternatives
anti-Rat (Rattus) MAP2K5 antibody for ELISA
Epitope
AA 380-430
33
30
15
15
15
14
14
11
11
9
6
6
6
5
4
4
3
2
2
2
2
1
1
1
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
168
88
81
11
7
7
6
4
3
3
3
3
2
2
2
2
1
1
Host
Rabbit
162
16
Clonality
Polyclonal
Conjugate
This MAP2K5 antibody is un-conjugated
9
9
9
5
5
5
5
5
5
5
5
5
5
4
4
4
4
Application
Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
110
63
47
28
24
22
4
3
2
1
1
1
Options
Supplier
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Available images

'Independent Validation' Badge
Lot Number 120828
Method validated Immunohistochemistry
Positive Control Human breast myoepithelial and glandular tissue
Negative Control Human breast adipose tissue
Notes Strong signal was detected in positive control sample, with minor background seen in negative control sample.
Primary Antibody
  • Antigen: Mitogen-Activated Protein Kinase Kinase 5 (MAP2K5)
  • Catalog number: ABIN754183
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 120828
Isotype Control Antibody
Secondary Antibody
  • Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
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  • Catalog number: Log in to see
  • Lot number: D07640BA
Controls
  • Positive control: Human breast myoepithelial and glandular tissue stained with antibody
  • Negative control: Human breast adipose tissue stained with antibody
  • Isotype control: Human breast myoepithelial and glandular tissue stained with isotype control
  • Secondary only control: Human breast myoepithelial and glandular tissue stained with secondary antibody only
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 (attached).
  • 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.
Validation Images
Immunohistochemistry validation image for anti-Mitogen-Activated Protein Kinase Kinase 5 (MAP2K5) (AA 380-430) antibody (ABIN754183) Figure 1: Human breast tissue stained with anti-MAP2K5 (brown) and counterstained wit...
Immunohistochemistry validation image for anti-Mitogen-Activated Protein Kinase Kinase 5 (MAP2K5) (AA 380-430) antibody (ABIN754183) Figure 2: Human breast adipose tissue stained with anti-MAP2K5 (brown) and countersta...
Immunohistochemistry validation image for anti-Mitogen-Activated Protein Kinase Kinase 5 (MAP2K5) (AA 380-430) antibody (ABIN754183) Figure 3: Human breast tissue stained with isotype control antibody (brown) and count...
Immunohistochemistry validation image for anti-Mitogen-Activated Protein Kinase Kinase 5 (MAP2K5) (AA 380-430) antibody (ABIN754183) Figure 4: Human breast tissue stained with secondary antibody only (brown) and counte...
Immunogen KLH conjugated synthetic peptide derived from human MEK5
Isotype IgG
Purification Purified by Protein A.
Alternative Name Mek5 (MAP2K5 Antibody Abstract)
Background

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. Plays an important role in the cascades of cellular responses evoked by changes in the environment. Mediates signaling for determination of cell fate such as differentiation and survival. Plays a crucial role in the apoptosis signal transduction pathway through mitochondria-dependent caspase activation. MAP3K5/ASK1 is required for the innate immune response, which is essential for host defense against a wide range of pathogens. Mediates signal transduction of various stressors like oxidative stress as well as by receptor-mediated inflammatory signals, such as the tumor necrosis factor (TNF) or lipopolysaccharide (LPS). Once activated, acts as an upstream activator of the MKK/JNK signal transduction cascade and the p38 MAPK signal transduction cascade through the phosphorylation and activation of several MAP kinase kinases like MAP2K4/SEK1, MAP2K3/MKK3, MAP2K6/MKK6 and MAP2K7/MKK7. These MAP2Ks in turn activate p38 MAPKs and c-jun N-terminal kinases (JNKs). Both p38 MAPK and JNKs control the transcription factors activator protein-1 (AP-1).

Subcellular location: Cytoplasm, Nucleus

Synonyms: MEK5, MAPKK5, Dual specificity mitogen-activated protein kinase kinase 5, MAPK/ERK kinase 5, MEK 5, MAP2K5, MAP kinase kinase 5

Gene ID 5607
UniProt Q13163
Research Area Cancer, Kinases/Phosphatases
Pathways MAPK Signaling, Neurotrophin Signaling Pathway
Application Notes WB 1:100-1000
IHC-P 1:100-500
IF(IHC-P) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage -20 °C
Storage Comment Store at -20°C
Expiry Date 12 months
Supplier Images
Immunohistochemistry (IHC) image for anti-MAP2K5 antibody (Mitogen-Activated Protein Kinase Kinase 5) (AA 380-430) (ABIN754183) Formalin-fixed and paraffin embedded rat brain tissue labeled with Anti-MEK5/MAP2K5 P...
Immunohistochemistry (IHC) image for anti-MAP2K5 antibody (Mitogen-Activated Protein Kinase Kinase 5) (AA 380-430) (ABIN754183) Independently Validated Antibody, image provided by Science Direct, badge number 0296...
Western Blotting (WB) image for anti-MAP2K5 antibody (Mitogen-Activated Protein Kinase Kinase 5) (AA 380-430) (ABIN754183) Mouse heart lysates probed with Rabbit Anti-MEK5 Antibody, Unconjugated at 1:5000 fo...
Product cited in: Chen, Hou, Ye, Chen, Cui, Tian, Li, Liu: "MicroRNA-143 regulates adipogenesis by modulating the MAP2K5-ERK5 signaling." in: Scientific reports, Vol. 4, pp. 3819, 2014 (PubMed).

Li, Dong, Wei, Wang, Zhang, Li: "Fatty acid synthase mediates the epithelial-mesenchymal transition of breast cancer cells." in: International journal of biological sciences, Vol. 10, Issue 2, pp. 171-80, 2014 (PubMed).