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Anti-KRT14 Antibody EP1612Y
Also for KRT14 (NM_000526)
|A synthetic peptide corresponding to residues near the C-terminus of human Cytokeratin 14 was used as an immunogen.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||WB: 1:20000; IP: 1:50; FC: 1:100; IHC-P: Use at an assay dependent concentration; ICC/IF: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Does not react with Mouse, Rat
|Homo sapiens keratin 14 (KRT14)|
|CK14; EBS3; EBS4; K14; NFJ|
|Keratins are cytoplasmic intermediate filament proteins expressed by epithelial cells. Cytokeratin 14 (CK-14) is a 50-kDa keratin expressed in abundance in human epidermal cells (1). During anagen, cell proliferation in the germinative matrix of the hair follicle gives rise to the fiber and inner root sheath. The transition from anagen to telogen is marked by downregulation of hair cortical specific keratins and the appearance of CK-14 in the epithelial sac to which the telogen hair fiber is anchored (2). A family with recessive epidermolysis bullosa simplex lacked expression of the basal cell keratin 14. The patients had severe generalized skin blistering that improved slightly with age. The basal cells of the patients did not express keratin 14 and contained no keratin intermediate filaments. The expression of keratin 5, the obligate copolymer of keratin 14, was slightly reduced (3). |
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Western blot - Cytokeratin 14 antibody [EP1612Y]; Anti-Cytokeratin 14 antibody [EP1612Y] at 1/20000 dilution + A431 cell lysate at 10 µg.Secondary.Goat anti-Rabbit HRP labeled at 1/2000 dilution.Predicted band size : 52 kDa.Observed band size : 48 kDa .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 14 antibody [EP1612Y]; ab51054 staining Cytokeratin 14 in human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer, pH 6.0. Samples were then permeabilized using 0.1% saponin/PBS, blocked with 4% BSA for 30 minutes at 25°C and then incubated with ab51054 at a 1/200 dilution for 16 hours at 4°C. The secondary used was a Texas Red conjugated goat anti-rabbit polyclonal used at a 1/100 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 14 antibody [EP1612Y]; ICC/IF image of stained A431cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody , 1/100 dilution) overnight at +4°C. The secondary antibody (green) was , DyLight? 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Flow Cytometry - Anti-Cytokeratin 14 antibody [EP1612Y]; Overlay histogram showing A431 cells stained with ab51054 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was was rabbit IgG (monoclonal) ( 1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.