Immunofluorescence analysis of EDG1 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 EDG1 Rabbit Polyclonal Antibody (PA1-1040) 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 localization in the membrane. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues S(359) H P Q K D D G D N P E T I(372) of mouse S1P1.|
|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|
|Immunohistochemistry (IHC)||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|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.
PA1-1040 detects S1P1 protein in transfected human and mouse cell samples. This antibody shows no cross-reactivity to S1P3.
PA1-1040 has successfully been used in immuno-fluorescence, immunohistochemistry, immunoprecipitation and Western blot procedures. By Western blot, this antibody detects an ~44 kDa protein representing recombinant S1P1 from transfected HEK293 cells. Immunofluorescence data demonstrates that S1P1 is localized to the plasma membrane after staining with PA1-1040.
The PA1-1040 immunogen is a synthetic peptide corresponding to residues S(359) H P Q K D D G D N P E T I(372) of mouse S1P1. This sequence is completely conserved in rat and 92% conserved in human S1P1. This peptide (Cat. # PEP-220) is available for use in neutralization and control experiments.
Sphingosine 1-phosphate (S1P) is bioactive lipid derived from the metabolism of sphingomyelin. S1P is predominately synthesized, stored, and released from platelets. When released, this molecule stimulates signaling pathways via G-protein coupled receptors. One such receptor is the heterotrimeric guanine nucleotide binding protein shingosine 1-phosphate receptor type 1 (S1P1,also known as EDG1).
Research has demonstrated that S1P regulates numerous cellular responses including, wound healing, cell growth, prevention of apoptosis, vascular maturation and cellular migration and survival. In studies involving S1P1 deficient embryos, blood vessels where incompletely maturated with smooth muscles cells, resulting in embryonic death. In contrast, a separate study demonstrated that overexpression of S1P1 led to the stimulation of cell migration.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Sphingosine 1-phosphate receptor 1 is required for retinal ganglion cell survival after optic nerve trauma.
PA1-1040 was used in immunocytochemistry, immunohistochemistry - frozen section, and western blot to assess the requirement for retinal ganglion cell survival after optic nerve trauma and sphingosine 1-phosphate receptor 1
|Joly S,Pernet V||Journal of neurochemistry (138:571)||2016|
Stress-induced production of chemokines by hair follicles regulates the trafficking of dendritic cells in skin.
PA1-1040 was used in immunohistochemistry to study the regulatory role of stress-induced chemokines on dendritic cell trafficking in skin
|Nagao K,Kobayashi T,Moro K,Ohyama M,Adachi T,Kitashima DY,Ueha S,Horiuchi K,Tanizaki H,Kabashima K,Kubo A,Cho YH,Clausen BE,Matsushima K,Suematsu M,Furtado GC,Lira SA,Farber JM,Udey MC,Amagai M||Nature immunology (13:744)||2012|
A polysaccharide, MDG-1, induces S1P1 and bFGF expression and augments survival and angiogenesis in the ischemic heart.
PA1-1040 was used in western blot to investigate the effect of the polysaccharide MDG-1 on specific gene expression and cell survival in ischemic hearts
|Wang S,Zhang Z,Lin X,Xu DS,Feng Y,Ding K||Glycobiology (20:473)||2010|