Human/Mouse/Rat MKK6 MAb (Clone 304328)

Name: Human/Mouse/Rat MKK6 MAb (Clone 304328)
Brand: R&D Systems
SKU: MAB1604

Catalog #: MAB1604 Datasheet / COA / SDS

Newer Version Available: AF1604

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Discontinued Product

MAB1604 has been discontinued and is replaced by AF16041. AF1604.

Detection of Human, Mouse, and Rat MKK6 by Western Blot.

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All MKK6 Antibodies

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Human/Mouse/Rat MKK6 MAb (Clone 304328) Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat MKK6 in Western blots. In Western blots, no cross-reactivity with recombinant human MKK3 is observed.

Source

Monoclonal Rat IgG_2B Clone # 304328

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human MKK6
Met1-Asp334
Accession # P52564.1

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

Immersion fixed human peripheral blood mononuclear cells

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

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Detection of Human, Mouse, and Rat MKK6 by Western Blot. Western blot shows lysates of B16-F1 mouse melanoma cell line and NRK rat normal kidney cell. PVDF membrane was probed with 0.5 µg/mL Rat Anti-Human/Mouse/Rat MKK6 Monoclonal Antibody (Catalog # MAB1604) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). For additional reference, recombinant human MKK3 and MKK6 (2 ng/lane) were included. A specific band for MKK6 was detected at approximately ??? kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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MKK6 in A431 Human Cell Line. MKK6 was detected in immersion fixed A431 human epithelial carcinoma cell line using Rat Anti-Human/Mouse/Rat MKK6 Monoclonal Antibody (Catalog # MAB1604) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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).

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