Immunofluorescence analysis of Cytokeratin 18 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin 18 (E431-1) Rabbit Monoclonal Antibody (MA514479) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide corresponding to residues of the human CK18|
|Storage buffer||tissue culture supernatant|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1:250|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-14479 targets Cytokeratin 18 in IHC (P) applications and shows reactivity with Human samples.
The MA5-14479 immunogen is a synthetic peptide corresponding to residues of the human CK18.
Cytokeratin 18, which belongs to the type A (acidic) subfamily of low molecular weight keratins, exists in combination with cytokeratin 8. Tissues from gastrointestinal tract, respiratory tract and urogenital tract, as well as endocrine and exocrine tissues and mesothelial cells are positive for cytokeratin 18. Typically, it is absent in squamous cell carcinoma.
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