Revision 1

#9384Store at -20C

1 Kit

(6 x 20 microliters)

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 Mol. Wt Isotype/Source
Keratin 17 (D73C7) Rabbit mAb 4543 20 µl 48 kDa Rabbit IgG
Keratin 8/18 (C51) Mouse mAb 4546 20 µl 46 Keratin 18. 55 Keratin 8. kDa Mouse IgG1
Keratin 18 (DC10) Mouse mAb 4548 20 µl 46 kDa Mouse IgG1
Keratin 19 (BA17) Mouse mAb 4558 20 µl 40 kDa Mouse IgG1
Pan-Keratin (C11) Mouse mAb 4545 20 µl 46-58 kDa Mouse IgG1
Keratin 7 (D1E4) XP® Rabbit mAb 4465 20 µl 52 kDa Rabbit IgG
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Cytokeratin Antibody Sampler Kit provides an economical means to evaluate the presence and status of selected keratin proteins. The kit provides enough primary and secondary antibodies to perform two Western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Keratins (cytokeratins) are intermediate filament proteins that are mainly expressed in epithelial cells. Keratin heterodimers composed of an acidic keratin (or type I keratin, keratins K9-K28) and a basic keratin (or type II keratin, keratins K1-K8 and K71-K80) assemble to form filaments. Keratin isoforms demonstrate tissue- and differentiation-specific profiles that make them useful as research and clinical biomarkers (1,2).

Dysregulation/mutations in keratin genes can lead to a variety of disorders affecting the skin, hair, nails, and other epithelial tissues (3). While expression of keratins can be variable, immunohistochemical staining of keratins is widely used to help in the identification and classification of epithelial tumors, and may also provide prognostic information.

Keratins 8 and 18 (K8/K18) are expressed in simple epithelia of normal tissue, as well as in adenocarcinomas of the breast, lung, ovary, and gastrointestinal tract. Keratin 17 is expressed in basal keratinocytes of stratified epithelia, hair follicles, and sebaceous glands. Onset of keratin 17 expression coincides with the definition of major epithelial lineages during skin development (4). Keratin 14 (K14) is expressed in basal cells of stratified epithelia, and in basal-like subtypes of breast cancer and squamous cell carcinomas. Keratin 19 (K19) is expressed in glandular epithelia, including the liver, gallbladder, and pancreas, as well as in adenocarcinomas of the breast, thyroid, and bile duct. Keratin 20 (K20) is expressed in gastrointestinal epithelium, urothelium, and Merkel cells in the skin, as well as in colorectal carcinomas and some urothelial carcinomas. Keratin 5/6 (K5/6) is expressed in basal cells of stratified epithelia, including the skin, prostate, and breast, as well as in basal-like breast cancers, squamous cell carcinomas, and some lung carcinomas. Keratin 7 (K7) is expressed in glandular epithelia, such as those in the lung, breast, and female reproductive tract, as well as in adenocarcinomas of the lung, breast, and ovary (5,6).

Keratins, particularly K8, K18, and K19, serve as biomarkers for identification of circulating tumor cells (CTCs) (5).

Post-translational modifications, including phosphorylation, acetylation, ubiquitylation, sumoylation, glycosylation, and transamidation, have been shown to affect the functions of keratins in normal and disease states (6). Understanding the molecular mechanisms underlying these PTMs may provide insights into cancer pathogenesis.

  1. Chang, L. and Goldman, R.D. (2004) Nat Rev Mol Cell Biol 5, 601-13.
  2. Schweizer, J. et al. (2006) J Cell Biol 174, 169-74.
  3. Sarma, A. (2022) Int J Biol Macromol 219, 395-413.
  4. McGowan, K.M. and Coulombe, P.A. (1998) J Cell Biol 143, 469-86.
  5. Werner, S. et al. (2020) Mol Aspects Med 72, 100817.
  6. Dmello, C. et al. (2019) J Biosci 44, 33.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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    Revision 1
    #9384

    Cytokeratin Antibody Sampler Kit

    Cytokeratin Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from HeLa and MCF7 cells using Keratin 7 (D1E4) XP® Rabbit mAb. Note that MCF7 cells are negative for Keratin 7, as expected.
    Cytokeratin Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from A-431, HeLa, and A549 cells using Keratin 17 (D73C7) Rabbit mAb. As expected, A549 cells are negative for keratin 17.
    Cytokeratin Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines, using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from HeLa and A431 cells, using Keratin 18 (DC10) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell types using Keratin 19 (BA17) Mouse mAb. As expected, the protein is absent in A-431 and HeLa cells.
    Cytokeratin Antibody Sampler Kit: Image 7 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Cytokeratin Antibody Sampler Kit: Image 8 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
    Cytokeratin Antibody Sampler Kit: Image 9 Expand Image
    Confocal immunofluorescent analysis of HeLa (left) and MCF7 (right) cells using Keratin 7 (D1E4) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cytokeratin Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of HeLa (left) and A549 (right) cells using Keratin 17 (D73C7) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cytokeratin Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma (bladder), using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic localization using Keratin 18 (DC10) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 14 Expand Image
    Immunhistochemical analysis of paraffin-embedded human breast carcinoma using Keratin 19 (BA17) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 15 Expand Image
    Flow cytometric analysis of Jurkat cells (blue) and HeLa cells (green) using Keratin 7 (D1E4) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    Cytokeratin Antibody Sampler Kit: Image 16 Expand Image
    Flow cytometric analysis of A549 cells (red) and HeLa cells (blue) using Keratin 17 (D73C7) Rabbit mAb.
    Cytokeratin Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Keratin 18 (DC10) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded MCF-7 cells (positive, left) and A-431 cells (negative, right), using Keratin 19 (BA17) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the lung, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 23 Expand Image
    Immunohistochemical analysis of paraffin-embedded human benign prostate hyperplasia, using Keratin 18 (DC10) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 26 Expand Image
    Confocal immunofluorescent analysis of HepG2 (left) and SH-SY5Y (right) cells using Keratin 18 (DC10) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Cytokeratin Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded H358 xenograft, using Pan-Keratin (C11) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded H358 xenograft, using Keratin 8/18 (C51) Mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 29 Expand Image
    Flow cytometric analysis of Jurkat cells (blue) and HT-29 cells (green) using Keratin 18 (DC10) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
    Cytokeratin Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Pan-Keratin (C11) mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 31 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Keratin 8/18 (C51) Mouse mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
    Cytokeratin Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Pan-Keratin (C11) mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 33 Expand Image
    Flow cytometric analysis of MCF-7 cells, using Keratin 8/18 (C51) Mouse mAb (blue) compared to a nonspecific negative control antibody (red).
    Cytokeratin Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Pan-Keratin (C11) mouse mAb.
    Cytokeratin Antibody Sampler Kit: Image 35 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Pan-Keratin (C11) Mouse mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
    Cytokeratin Antibody Sampler Kit: Image 36 Expand Image
    Flow cytometric analysis of A-431 cells using Pan-Keratin (C11) Mouse mAb (solid line) versus a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed line). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.