Immunofluorescence analysis of MUC5B was performed using 70 % confluent log phase A549 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 MUC5B (5B19-2E) Mouse Monoclonal antibody (377400) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at 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 punctated cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Synthetic peptide derived from the N-terminal region of the human MUC5B protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
Mucins (MUCs) are heavily glycosylated glycoproteins that are synthesized either as membrane-bound or secreted proteins. Mucins are noteworthy because they have an aberrant expression profile in various malignancies. Mucins normally play a protective role for epithelial tissues and are expressed in the gastrointestinal, respiratory, and genito-urinary tracts. Mucins may also be involved in the renewal and differentiation of the epithelium, and the modulation of cell adhesion and cell signaling. MUC5B is thought to play a vital role in maintaining oral health, and may also be involved in respiratory and oral disease.2 MUC5B expression has been studied in gastric carcinoma as well as chronic obstructive pulmonary disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.