Revision 1

#18625Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P42771

Entrez-Gene Id:

#1029

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
p16 INK4A Rabbit Capture mAb 38991 1 ea Green (Lyophilized)
p16 INK4A Rabbit HRP-linked mAb 89398 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 #18625 Positive Control Type 2 15550 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 p16 INK4A ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p16 INK4A. 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 p16 INK4A 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 p16 INK4A.

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

IMPORTANT: This FastScan ELISA Kit requires 4 washes at Step 6 of the protocol.

Specificity/Sensitivity

The FastScan p16 INK4A ELISA Kit detects endogenous levels of p16 INK4A, 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

Members of the INK4 family of cyclin-dependent kinase inhibitors include p16 INK4A, p15 INK4B, p18 INK4C, and p19 INK4D. The INK4 family members inhibit cyclin-dependent kinases 4 and 6 (CDK4 and CDK6), causing cell cycle arrest in G1 phase. The INK4A-ARF-INK4B locus on chromosome 9p21, frequently lost in human cancer, encodes the INK4 family members p16 INK5A and p15 INK4B, as well as the unrelated protein, ARF (1).

p16 INK4A expression, typically repressed in the absence of stress, is thought to drive cells into senescence, and p16 INK4A expression is a commonly used marker of senescent cells (2,3). p16 INK4A protein expression is often altered in human cancer, and high expression is currently used as a predictive biomarker in cervical cancer (3-5).

  1. Kim, W.Y. and Sharpless, N.E. (2006) Cell 127, 265-75.
  2. LaPak, K.M. and Burd, C.E. (2014) Mol Cancer Res 12, 167-83.
  3. Romagosa, C. et al. (2011) Oncogene 30, 2087-97.
  4. Ishikawa, M. et al. (2006) Int J Gynecol Cancer 16, 347-53.
  5. Queiroz, C. et al. (2006) Pathol Res Pract 202, 77-83.

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 1
    #18625

    FastScan p16 INK4A ELISA Kit

    FastScan™ p16 INK4A ELISA Kit: Image 1 Expand Image
    Figure 1. p16 INK4A protein is expressed in HeLa cells but absent in MCF7 cells. The relationship between lysate protein concentration from MCF7 and HeLa cells and the absorbance at 450 nm using the FastScan p16 INK4A ELISA Kit #18625 is shown in the upper figure. The corresponding western blots using p16 INK4A antibody (left panel) and GAPDH antibody (right panel) are shown in the lower figure. Untreated MCF7 or untreated HeLa cells (90% confluence) were collected and then lysed.