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Immunofluorescence analysis of Thomsen Friedenreich Antigen was performed using 70% confluent log phase HT-29 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 Thomsen-Friedenreich Antigen (A78-G/A7) Mouse Monoclonal Antibody (MA513466) at 2µg/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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Pig|
|Host / Isotype||Mouse / IgM, kappa|
|Immunogen||Neuraminidase-treated human red blood cells|
|Purification||Ammonium sulfate precipitation|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|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|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:25-1:50|
* 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 1 publications below|
MA5-13466 targets Thomsen-Friedenreich Antigen in immunofluorescence and immohistochemistry (Paraffin) applications and shows reactivity with Human and Rat samples. Staining of formalin-fixed tissues requires boiling tissue section in 10 mM citrate buffer, pH 6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.
The MA5-13466 immunogen is neuraminidase-treated human red blood cells.
The Thomsen-Fridenreich antigen acts as an oncofetal antigen, with low expression in normal adult tissues but increasing to fetal levels of expression in hyperplasia or malignancy. It is considered as a pan-carcinoma marker.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Infection of swine ex vivo tissues with avian viruses including H7N9 and correlation with glycomic analysis.
MA5-13466 was used in immunohistochemistry to study the sialylated N-glycan and O-glycan content of the pig resiratory tract and the significance for ex vivo infection by avian viruses
|Chan RW,Karamanska R,Van Poucke S,Van Reeth K,Chan IW,Chan MC,Dell A,Peiris JS,Haslam SM,Guan Y,Nicholls JM||Influenza and other respiratory viruses (7:1269)||2013|