Immunofluorescence analysis of Arfaptin 2 was performed using 70% confluent log phase MDA-MB-231 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 Arfaptin 2 Rabbit Polyclonal Antibody (Product # 40-2400) at 2µg/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 (Product # 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 (Product # 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 cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the N-terminal region of the human Arfaptin 2 protein (ADP-ribosylation factor interacting protein 2, AFP2, Partner of Rac1, or POR1)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2 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.
|Immunofluorescence (IF)||See 2 publications below|
Putative target protein of ADP-ribosylation factor. Involved in membrane ruffling.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||Structural basis for membrane binding specificity of the Bin/Amphiphysin/Rvs (BAR) domain of Arfaptin-2 determined by Arl1 GTPase.||Nakamura K,Man Z,Xie Y,Hanai A,Makyio H,Kawasaki M,Kato R,Shin HW,Nakayama K,Wakatsuki S||The Journal of biological chemistry (287:25478)||2012|
|Human||Not Cited||Arfaptins are localized to the trans-Golgi by interaction with Arl1, but not Arfs.||Man Z,Kondo Y,Koga H,Umino H,Nakayama K,Shin HW||The Journal of biological chemistry (286:11569)||2011|