Immunofluorescence analysis of Dog1 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 Dog1 (SP31) Rabbit Monoclonal Antibody (MA5-14582) at 2 µg/ml 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 Alexa Fluor® 555 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, Rat|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide of human Dog1|
|Storage buffer||tissue culture supernatant|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:25-50|
|Western Blot (WB)||2-4 µg/ml|
* 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.
|Immunohistochemistry (IHC)||See 4 publications below|
MA5-14582 targets Dog1 in IHC (P) applications and shows reactivity with Human samples.
The MA5-14582 immunogen is synthetic peptide of human Dog1.
DOG1 is a cell surface protein of unknown function selectively expressed in GIST. Among GIST cases with KIT mutations the DOG1 antibody identified 11% more cases than c-Kit antibody. DOG1 identifies vast majority of both c-Kit negative and PDGFRA mutated GIST cases that may still benefit from imatinib mesylate (Gleevac), an inhibitor of the kit tyrosin kinase. In addition, DOG1 immunoreactivity is seen in fewer cases of mesanchymal, epithelial tumors and melanomas when compared with c-Kit. The use of this highly sensitive and specific novel marker will increase the accuracy of GIST diagnosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
TMEM16A/ANO1 is differentially expressed in HPV-negative versus HPV-positive head and neck squamous cell carcinoma through promoter methylation.
MA5-14582 was used in immunohistochemistry to elucidate differential expression of TMEM16A/ANO1 in HPV-positive and HPV-negative head and neck squamous cell carcinoma through promoter methylation
|Dixit R,Kemp C,Kulich S,Seethala R,Chiosea S,Ling S,Ha PK,Duvvuri U||Scientific reports (5:null)||2015|
Plexiform fibromyxoma (plexiform angiomyxoid myofibroblastic tumor) of stomach: an unusual presentation as a fistulating abscess.
MA5-14582 was used in immunohistochemistry to report on a case of plexiform fibromyxoma of the stomach.
|Lee PW,Yau DT,Lau PP,Chan JK||International journal of surgical pathology (22:286)||2014|
Contribution of DOG1 expression to the diagnosis of gastrointestinal stromal tumors.
MA5-14582 was used in immunohistochemistry to study the diagnostic value of the immunohistochemical expression of DOG1 in gastrointestinal stromal tumors
|Kara T,Serinsoz E,Arpaci RB,Gubur O,Orekici G,Ata A,Colak T,Arican A||Pathology, research and practice (209:413)||2013|
The KIT Exon 11 Stop Codon Mutation in Gastrointestinal Stromal Tumors: What Is the Clinical Meaning?
MA5-14582 was used in immunohistochemistry to study the potential clinical significance in gastrointestinal stromal tumors of KIT exon 11 stop codon mutations
|Michelucci A,Chiappetta C,Cacciotti J,Veccia N,Astri E,Leopizzi M,Prosperi Porta R,Petrozza V,Della Rocca C,Bevilacqua G,Cavazzana A,Di Cristofano C||Gut and liver (7:35)||2013|