Cytokeratin 14 Mouse Monoclonal Antibody [A2C11]
Catalog# EM1901-32
Cytokeratin 14 Mouse Monoclonal Antibody [A2C11]
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WB
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IHC-P
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IF-Cell
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Human
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unconjugated
Overview
Product Name
Cytokeratin 14 Mouse Monoclonal Antibody [A2C11]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Synthetic peptide within Human Cytokeratin 14 aa 423-472 / 472.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell
Molecular Weight
Predicted band size: 52 kDa
Positive Control
A431 cell lysate, A431, human lung squamous carcinoma tissue, human skin tissue, human tonsil tissue.
Conjugation
unconjugated
Clone Number
A2C11
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein G affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IHC-P
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1:1:1,000
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IF-Cell
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1:100
Target
Function
This gene encodes a member of the keratin family, the most diverse group of intermediate filaments. This gene product, a type I keratin, is usually found as a heterotetramer with two keratin 5 molecules, a type II keratin. Together they form the cytoskeleton of epithelial cells. Mutations in the genes for these keratins are associated with epidermolysis bullosa simplex. The nonhelical tail domain is involved in promoting KRT5-KRT14 filaments to self-organize into large bundles and enhances the mechanical properties involved in resilience of keratin intermediate filaments in vitro. Expressed in the corneal epithelium (at protein level). Detected in the basal layer, lowered within the more apically located layers specifically in the stratum spinosum, stratum granulosum but is not detected in stratum corneum. Strongly expressed in the outer root sheath of anagen follicles but not in the germinative matrix, inner root sheath or hair. A form of epidermolysis bullosa simplex, a group of skin fragility disorders characterized by skin blistering due to cleavage within the basal layer of keratinocytes, and erosions caused by minor mechanical trauma. There is a broad spectrum of clinical severity ranging from minor blistering on the feet, to subtypes with extracutaneous involvement and a lethal outcome. EBS1A is an autosomal dominant form characterized by generalized intraepidermal skin blistering that begins and is very prominent at birth. EBS1A may be life-threatening in the first year of life. Tendency to blistering diminishes in adolescence.
Background References
1. Bousquet O. et al. The nonhelical tail domain of keratin 14 promotes filament bundling and enhances the mechanical properties of keratin intermediate filaments in vitro. J. Cell Biol. 155:747-754(2001).
2. Schweizer J et al. "New consensus nomenclature for mammalian keratins". The Journal of Cell Biology. 174 (2): 169–74(2006).
Sequence Similarity
Belongs to the intermediate filament family.
Tissue Specificity
Expressed in the corneal epithelium (at protein level). Detected in the basal layer, lowered within the more apically located layers specifically in the stratum spinosum, stratum granulosum but is not detected in stratum corneum. Strongly expressed in the outer root sheath of anagen follicles but not in the germinative matrix, inner root sheath or hair. Found in keratinocytes surrounding the club hair during telogen.
Post-translational Modification
A disulfide bond is formed between rather than within filaments and promotes the formation of a keratin filament cage around the nucleus.; Ubiquitinated by the BCR(KLHL24) E3 ubiquitin ligase complex.
Subcellular Location
Cytoplasm, Nucleus.
UNIPROT
Synonyms
CK 14 antibody
CK-14 antibody
ck14 antibody
Cytokeratin 14 antibody
Cytokeratin-14 antibody
Cytokeratin14 antibody
Dowling Meara antibody
EBS3 antibody
EBS4 antibody
Epidermolysis bullosa simplex antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of Cytokeratin 14 on different lysates with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 dilution.
Lane 1: A431 cell lysate
Lane 2: MCF7 cell lysate (negative)
Lane 3: PANC-1 cell lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 1 minute 59 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-32) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Relative expression (RE)
Immunocytochemistry analysis of A431 (positive) and PANC-1 (negative) labeling Cytokeratin 14 with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human lung squamous carcinoma tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-32) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human skin tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-32) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-32) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"