Immunofluorescence analysis of APS was performed using 70% confluent log phase Neuro-2a 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 APS Rabbit Polyclonal Antibody (PA1-1062) 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 cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues E(605) A T L G R A R A V E N Q Y S(619) from mouse APS.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|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)||1:500|
* 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.
PA1-1062 detects APS from mouse samples.
PA1-1062 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~ 84 kDa protein corresponding to APS from recombinant mouse protein and 3T3L1 extracts.
The PA1-1062 immunogen is a synthetic peptide corresponding to residues E(605)A T L G R A R A V E N Q Y S(619) from mouse APS. This peptide (Cat. # PEP-246) can be used in neutralization and control experiments.
APS (adapter protein with Pleckstrin homology and Src homology 2 domains) a member of the Lnk family, is an adaptor protein that is involved in B-cell signaling, insulin signaling, cytoskeletal reorganization. It is tyrosine phosphorylated by JAK2, KIT and other kinases during B-cell receptor stimulation of many different cytokines, chemokines and leukokines. APS has been shown to inhibit the JAK-STAT pathway in collaboration with c-Cbl. It is located in the cytoplasm in pre-PDGF (platelet derived growth factor) stimulated cells and localizes to the plasma membrane and peripheral regions upon PDGF stimulation.
This protein is expressed in B-cells and mast cells when detected in haematopoietic cell lines yet is also expressed in many different tissues including spleen, prostrate, testis, uterus, small intestine and skeletal muscle.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.