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Key Product Details

Validated by

Orthogonal Validation

Species Reactivity

Human, Mouse, Rat, Bovine, Canine, Chicken, Monkey, Rabbit, Reptile, Zebrafish

Applications

CyTOF-reported, Dual RNAscope ISH-IHC, Flow (Intracellular), Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Single Cell Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Kappa/IgG1 Kappa Clone # AE-1/AE-3

Concentration

0.2 mg/ml

Product Summary for Cytokeratin, pan Antibody (AE-1/AE-3)

Immunogen

Human epidermal keratin

Reactivity Notes

Reptile reactivity reported in scientific literature (PMID: 11351328). Zebrafish reactivity reported in scientific literature (PMID: 30970016).

Localization

Cytoplasmic

Specificity

Twenty human keratins are resolved with two-dimensional gel electrophoresis into acidic (pI 6.0) subfamilies. This antibody cocktail recognizes acidic (Type I or LMW) and basic (Type II or HMW) cytokeratins, which 67kDa (CK1); 64kDa (CK3); 59kDa (CK4); 58kDa (CK5); 56kDa (CK6); 52kDa (CK8); 56.5kDa (CK10); 50kDa (CK14); 50kDa (CK15); 48kDa (CK16); 40kDa (CK19). Many studies have shown the usefulness of keratins as markers in cancer research and tumor diagnosis. AE-1/AE-3 is a broad spectrum anti pan-cytokeratin antibody cocktail, which differentiates epithelial tumors from non-epithelial tumors e.g. squamous vs. adenocarcinoma of the lung, liver carcinoma, breast cancer, and esophageal cancer. It has been used to characterize the source of various neoplasms and to study the distribution of cytokeratin containing cells in epithelia during normal development and during the development of epithelial neoplasms. This antibody stains cytokeratins present in normal and abnormal human tissues and has shown high sensitivity in the recognition of epithelial cells and carcinomas.

Marker

Epithelial Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 Kappa/IgG1 Kappa

Description

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-33200)

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C. Antibody is stable for 24 months. Non-hazardous.

Scientific Data Images for Cytokeratin, pan Antibody (AE-1/AE-3)

Detection of Cytokeratin, pan in Human Tissue in Dual RNAscope ISH-IHC

Detection of Cytokeratin, pan in Human Tissue in Dual RNAscope ISH-IHC

FFPE tissue sections of human metastatic tonsil were probed for Pan Cytokeratin mRNA (ACD RNAScope probe, catalog # 310221; Fast Red chromogen, ACD catalog # 322500). Adjacent tissue section was processes for IHC using mouse monoclonal antibody (Novus catalog # NBP2-29429) at 0.3 ug/mL for 1 hour at room temperature followed by incubation with the anti-mouse IgG VisUCyte HRP Polymer Antibody (Novus Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue).
Staining of Cytokeratin, pan in MCF-7 Cells Using Conjugated Cytokeratin, pan Antibody

Staining of Cytokeratin, pan in MCF-7 Cells Using Conjugated Cytokeratin, pan Antibody

MCF-7 cells stained with Biotin conjugated version of pan Cytokeratin antibody and SAv-A488. A) SAv-A488 only at a dilution of 1:500, B) pan CK at a dilution of 1:200, C) pan CK at a dilution of 1:400, D) pan CK at a dilution of 1:800. ICC/IF image submitted by a verified customer review.
Immunohistochemical Staining of Cytokeratin, pan in Paraffin Embedded Human Colon Carcinoma

Immunohistochemical Staining of Cytokeratin, pan in Paraffin Embedded Human Colon Carcinoma

Analysis using Azide and BSA Free version of NBP2-29429. Human Colon Carcinoma stained with pan Cytokeratin Monoclonal Antibody cocktail (AE-1/AE3).

Applications for Cytokeratin, pan Antibody (AE-1/AE-3)

Application
Recommended Usage

Flow Cytometry

1-2 ug/million cells

Immunocytochemistry/ Immunofluorescence

1-2 ug/ml

Immunohistochemistry-Frozen

0.5-1.0ug/ml

Immunohistochemistry-Paraffin

0.25-0.5 ug/ml

Single Cell Western

1:10

Western Blot

1-2 ug/ml
Application Notes
Use in ICC/IF reported in scientific literature (PMID:34376789)
Immunohistochemistry (Formalin-fixed): 0.25-0.5ug/ml for 30 min at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.
Western Blot: 1-2ug/ml for 2 hours at RT.
The staining pattern of the pan cytokeratin antibody cocktail may be different than that of either antibody separately.
This antibody cocktail recognizes acidic (Type I or LMW) and basic (Type II or HMW) cytokeratins, which 67 kDa (CK1) ; 64 kDa (CK3) ; 59 kDa (CK4) ; 58 kDa (CK5) ; 56 kDa (CK6) ; 52 kDa (CK8) ; 56.5 kDa (CK10) ; 50k Da (CK14) ; 50 kDa (CK15) ; 48 kDa (CK16) ; 40 kDa (CK19) . The pan cytokeratin cocktail does not react with keratin 18, which is also expressed in carcinomas. As such, negative staining with NBP2-29429 in of itself may not be sufficient evidence to rule out the possibility of a carcinoma (Ordonez, 2013) .
For example, hepatocellular, adrenal cortical, clear cell renal and chromophobe renal cell carcinomas have been reported to be negative for the pan cytokeratin antibody. In this regard, the pan cytokeratin antibody can be used as part of a screening panel to more extensively define the tumor cell lineages.
The pan cytokeratin antibody may cross-react with GFAP, leading to aberrant positive staining of glial tumors such as ependymoma, glioblastoma, or schwannoma (Ordonez, 2013) . Use in Immunohistochemistry reported in scientific literature (PMID: 29169625) . This Cytokeratin, pan Antibody (AE-1/AE-3) is validated for CyTOF from a verified customer review.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using NBP2-29429 in the following applications:

Published Applications

Read 54 publications using NBP2-29429 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS with 0.05% BSA

Preservative

0.05% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C.

Background: Cytokeratin, pan

Cytokeratins are a family of intermediate filamentous proteins that are expressed by epithelial cells (1,2). Cytokeratins range in size with a theoretic molecular weight varying from approximately 40 kDa to 68 kDa (2,3). The cytokeratin family consists of 20 polypeptides that are further divided into two main groups based on isoelectric point and molecular weight (1-3). The type I group are smaller, acidic polypeptides designated as cytokeratin 9 through cytokeratin 20 (CK9 - CK20) (1-4). Conversely, CK1 - CK8 belong to the type II group, classified as larger, basic or neutral polypeptides (1-4). Structurally, cytokeratins have homologous basic structure with other intermediate filaments; they possess a 300-315 amino acid (aa) central helical region that consists of four conserved domains (1A, 2A, 1B, and 2B) which are separated by linker domains (L1, L12, and L2) (1,5). Additionally, flanking this central region, both the amino- and carboxyl-terminal ends have a homologous subdomain (H), a variable domain (V), and charged end subdomains (E) (1). Furthermore, the central rod of one cytokeratin monomer binds with another monomer to form a coiled-coil dimer which subsequently binds another dimer to form a tertramer (3). Finally, many tetramers join together to ultimately form an intermediate filament of approximately 10nm in diameter (1-3, 5). Cytokeratins are expressed as pairs, typically with a type I and type II member; for example, CK10 pairs with CK1 (1,3).

Epithelial cells express multiple subtypes of cytokeratins which can be used to classify epithelial cell type or differentiation status, as well tumor progression or diagnosis (2). Cytokeratins are important for both stability and integrity of epithelial cells and function in intracellular signaling, from wound healing to apoptosis (1). Cytokeratins are useful immunohistochemistry tumor markers and antibodies to cytokeratins are a common pathological tool (1,3,6). Cytokeratin pan antibody is an antibody cocktail mixture that can detect multiple cytokeratins and reacts to multiple epithelial tissues (1,3,6). For example, AE-1/AE-3 is a commonly used specific pan cytokeratin that detects cytokeratins 1-8, 10, 14-16 and 19 (1,3,6).

Given the role of cytokeratins in the structural integrity of epithelial cells, mutations in cytokeratins have been shown to play a role in a variety of human diseases including epidermolysis bullosa simplex (EBS) (4,5). EBS is an autosomal dominant disorder that is caused by missense mutations in either CK5 or CK14 (5). Other known cytokeratin-related disorders include bullous ichthyosis, a skin disorder characterized by redness, blistering, and hyperkeratosis, and epidermolytic palmoplantar keratoderma (EPPK), which results in hyperkeratosis on the palms and soles of the body (7).

References

1. Awasthi, P., Thahriani, A., Bhattacharya, A., Awasthi, P., & Keratins, B. A. (2016). Keratins or cytokeratins: a review article. Journal of Advanced Medical and Dental Sciences Research. https://10.21276/jamdsr.2016.4.4.30

2. Southgate, J., Harnden, P., & Trejdosiewicz, L. K. (1999). Cytokeratin expression patterns in normal and malignant urothelium: a review of the biological and diagnostic implications. Histology and histopathology. https://doi.org/10.14670/HH-14.657

3. Belaldavar, C., Mane, D. R., Hallikerimath, S., Kale, A. D. (2016). Cytokeratins: Its role and expression profile in oral health and disease. Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology. https://doi.org/10.1016/j.ajoms.2015.08.001.

4. Linder S. (2007). Cytokeratin markers come of age. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine. https://doi.org/10.1159/000107582

5. Jacob, J. T., Coulombe, P. A., Kwan, R., & Omary, M. B. (2018). Types I and II Keratin Intermediate Filaments. Cold Spring Harbor perspectives in biology. https://doi.org/10.1101/cshperspect.a018275

6. Ordonez N. G. (2013). Broad-spectrum immunohistochemical epithelial markers: a review. Human pathology. https://doi.org/10.1016/j.humpath.2012.11.016

7. McLean, W. H., & Moore, C. B. (2011). Keratin disorders: from gene to therapy. Human molecular genetics. https://doi.org/10.1093/hmg/ddr379

Alternate Names

AEI2;CK1;EHK;EHK1;EPPK;K1;Keratin, type II cytoskeletal 1;KRT1A;NEPPK

Gene Symbol

KRT1

Product Documents for Cytokeratin, pan Antibody (AE-1/AE-3)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Cytokeratin, pan Antibody (AE-1/AE-3)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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FAQs for Cytokeratin, pan Antibody (AE-1/AE-3)

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  • Q: Could you tell us recommended antigen retrieval method for this antibody?-EndFragment-->

    A: Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes. Optimal dilution of antibody for IHC should be determined by the user. 

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