Human/Mouse/Rat MKK6 Antibody

Catalog # Availability Size / Price Qty
AF1604
AF1604-SP
Detection of Human/Mouse/Rat MKK6 by Western Blot.
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Human/Mouse/Rat MKK6 Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat MKK6 in Western blots.
Source
Polyclonal Rabbit IgG
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.05 µg/mL
See below
Knockout Validated
MKK6 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in MKK6 knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human/Mouse/Rat MKK6 antibody by Western Blot. View Larger

Detection of Human/Mouse/Rat MKK6 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, C2C12 mouse myoblast cell line, TS1 mouse helper T cell line, and B16-F1 mouse melanoma cell line. PVDF membrane was probed with 0.05 µg/mL Rabbit Anti-Human/Mouse/Rat MKK6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1604) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). For additional reference, recombinant human MKK6 (2 ng) was included. A specific band for MKK6 was detected at approximately 38 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Knockout Validated Western Blot Shows Human MKK6 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human MKK6 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and MKK6 knockout HeLa cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human/Mouse/Rat MKK6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1604) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for MKK6 at approximately 39 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: MKK6

Mitogen-activated protein kinase kinase 6 (MKK6), also known as MEK6, and the closely related MKK3 are activated by pro-inflammatory cytokines and environmental stresses (1, 2). Both MKK3 and MKK6 are dual specificity protein kinases, phosphorylating and activating the p38 MAP kinases at Thr and Tyr positions within the phosphoacceptor sequence Thr-Gly-Tyr (3). Activation of MKK6 occurs through phosphorylation at Ser207 and Thr211 (4) by several upstream MAPK kinase kinases, including ASK1 and TAK1 (5, 6). Purification of E. coli-expressed wild-type active MKK6 capable of p38 alpha phosphorylation has been previously noted (7).

References
  1. Goedert, M. et al. (1997) EMBO J. 16:3563.
  2. Moriguchi, T. et al. (1996) J. Biol. Chem. 271:26981.
  3. Raingeaud, J. et al. (1995) J. Biol. Chem. 270:7420.
  4. Han, J. et al. (1996) J. Biol. Chem. 271:2886.
  5. Ichijo, H. et al. (1997) Science 275:90.
  6. Moriguchi, T. et al. (1996) J. Biol. Chem. 271:13675.
  7. Keesler, G. et al. (1998) Protein Expr. Purif. 14:221.
Long Name
Mitogen-activated Protein Kinase Kinase 6
Entrez Gene IDs
5608 (Human); 26399 (Mouse); 114495 (Rat)
Alternate Names
MAP kinase kinase 6; MAP2K6; MAPK/ERK kinase 6; MAPKK6dual specificity mitogen-activated protein kinase kinase 6; MEK6; MEK6MAPKK 6; mitogen-activated protein kinase kinase 6; MKK6; MKK6MEK 6; PRKMK6; PRKMK6protein kinase, mitogen-activated, kinase 6 (MAP kinase kinase 6); SAPKK3; SAPKK3EC 2.7.12.2

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Product Specific Notices

The purchase of this product conveys to the buyer the limited, non-exclusive, non-transferable right (without the right to resell, repackage, or further sublicense) to use these reagents for research purposes only. No other license is granted to the buyer whether expressly, by implication, by estoppel or otherwise. In particular, the purchase of this product does not include nor carry any right or license to use, develop, or otherwise exploit this product commercially, which includes without limitation, provision of services to a third party, generation of commercial databases, clinical diagnostics or therapeutics, or drug development. This product is sold pursuant to a license from the University of Massachusetts, and the University of Massachusetts reserves all other rights to use these reagents. For information on purchasing a license for uses other than research, please contact the University of Massachusetts, Office of Technology Management, 333 South Street, Suite 400, Shrewsbury, Massachusetts, 01545.

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