Revision 6

#46948Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H R

UniProt ID:

#Q9UHD2

Entrez-Gene Id:

#29110

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color
FastScan ELISA Microwell Strip Plate, 96 Well 53257 96 tests
Phospho-TBK1/NAK (Ser172) Rabbit Capture mAb 59777 1 ea Green (Lyophilized)
TBK1/NAK Rabbit HRP-linked mAb 74257 1 ea Red (Lyophilized)
FastScan ELISA Capture Antibody Diluent 16076 3 ml Green
FastScan ELISA HRP Antibody Diluent 28120 3 ml
TMB Substrate 7004 11 ml
STOP Solution 7002 11 ml
Sealing Tape 54503 1 ea
ELISA Wash Buffer (20X) 9801 25 ml
FastScan ELISA Cell Extraction Buffer (5X) 69905 10 ml
FastScan ELISA Cell Extraction Enhancer Solution (50X) 25243 1 ml
FastScan ELISA Kit #46948 Positive Control Type 2 89559 1 ea

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The FastScan Phospho-TBK1/NAK (Ser172) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of TBK1/NAK when phosphorylated at Ser172. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-TBK1/NAK (Ser172) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-TBK1/NAK (Ser172).

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The FastScan Phospho-TBK1/NAK (Ser172) ELISA Kit detects endogenous levels of TBK1/NAK when phosphorylated at Ser172, as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

TBK1 (TANK-binding kinase 1)/NAK (NF-κB activating kinase) is an IκB kinase (IKK)-activating kinase and can activate IKK through direct phosphorylation (1). TBK1 was identified through association with the TRAF binding protein, TANK, and found to function upstream of NIK and IKK in the activation of NF-κB (2). TBK1 induces IκB degradation and NF-κB activity through IKKβ. TBK1 may mediate IKK and NF-κB activation in response to growth factors that stimulate PKCε activity (1). TBK1 plays a pivotal role in the activation of IRF3 in the innate immune response (3).

  1. Tojima, Y. et al. (2000) Nature 404, 778-82.
  2. Pomerantz, J.L. and Baltimore, D. (1999) EMBO J 18, 6694-704.
  3. Fitzgerald, K.A. et al. (2003) Nat Immunol 4, 491-6.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 6
    #46948

    FastScan Phospho-TBK1/NAK (Ser172) ELISA Kit

    FastScan™ Phospho-TBK1/NAK (Ser172) ELISA Kit: Image 1 Expand Image
    Figure 1. Treatment of differentiated THP-1 cells with LPS stimulates phosphorylation of TBK1/NAK at Ser172, but does not affect the level of total TBK1/NAK. The relationship between lysate protein concentration from TPA differentiated THP-1 cells, either untreated or LPS-treated, and the absorbance at 450 nm using the FastScan Phospho-TBK1/NAK (Ser172) ELISA Kit #46948 is shown in the upper figure. The corresponding western blots using phospho-TBK1/NAK (Ser172) antibody (left panel) and TBK1/NAK antibody (right panel) are shown in the lower figure. THP-1 cells were differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (200 nM, 4 d), allowed to rest in fresh growth media for 3 days, then treated with Lipopolysaccharides (LPS) #14011 (1.0 μg/mL, 1 hr).