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Fas Ligand Rabbit pAb

Blank control: Mouse Kidney (blue). Primary Antibody:Rabbit Anti-phospho-Fas Ligand antibody (AP52211,Green); Dilution: 1 µg in 100 µL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37℃. Primary antibody (AP52211, 1 µg /1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min on ice. Acquisition of 20,000 events was performed.
Sample: Bone (mouse) Lysate at 40 ug
Primary: Anti- Fas ligand (AP52211) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kD
Observed band size: 46 kD
Tissue/cell: human rectal carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-FasL Polyclonal Antibody, Unconjugated(AP52211) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(AP52211-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Fas Ligand) Polyclonal Antibody, Unconjugated (AP52211) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IHC-F, IF
Primary Accession	P48023
Reactivity	Rat, Human, Mouse
Host	Rabbit
Clonality	Polyclonal
Calculated MW	31485 Da
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from human Fas Ligand
Epitope Specificity	196-281/281
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Cell membrane; Single-pass type II membrane protein. Secreted. Cytoplasmic vesicle lumen. Lysosome lumen. Note=May be released into the extracellular fluid, probably by cleavage form the cell surface. Is internalized into multivesicular bodies of secretory lysosomes after phosphorylation by FGR and monoubiquitination.
DISEASE	Defects in FASLG are the cause of autoimmune lymphoproliferative syndrome type 1B (ALPS1B) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	This gene is a member of the tumor necrosis factor superfamily. The primary function of the encoded transmembrane protein is the induction of apoptosis triggered by binding to FAS. The FAS/FASLG signaling pathway is essential for immune system regulation, including activation-induced cell death (AICD) of T cells and cytotoxic T lymphocyte induced cell death. It has also been implicated in the progression of several cancers. Defects in this gene may be related to some cases of systemic lupus erythematosus (SLE). Alternatively spliced transcript variants have been described. [provided by RefSeq, Nov 2014]

Additional Information
Gene ID	356
Other Names	Tumor necrosis factor ligand superfamily member 6, Apoptosis antigen ligand, APTL, CD95 ligand, CD95-L, Fas antigen ligand, Fas ligand, FasL, CD178, Tumor necrosis factor ligand superfamily member 6, membrane form, Tumor necrosis factor ligand superfamily member 6, soluble form, Receptor-binding FasL ectodomain, Soluble Fas ligand, sFasL, ADAM10-processed FasL form, APL, FasL intracellular domain, FasL ICD, SPPL2A-processed FasL form, SPA, FASLG, APT1LG1, CD95L, FASL, TNFSF6
Dilution	WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1 µg/Test
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	FASLG
Synonyms	APT1LG1, CD95L, FASL, TNFSF6
Function	Cytokine that binds to TNFRSF6/FAS, a receptor that transduces the apoptotic signal into cells (PubMed:26334989, PubMed:9228058). Involved in cytotoxic T-cell-mediated apoptosis, natural killer cell-mediated apoptosis and in T-cell development (PubMed:7528780, PubMed:9228058, PubMed:9427603). Initiates fratricidal/suicidal activation-induced cell death (AICD) in antigen- activated T-cells contributing to the termination of immune responses (By similarity). TNFRSF6/FAS-mediated apoptosis also has a role in the induction of peripheral tolerance (By similarity). Binds to TNFRSF6B/DcR3, a decoy receptor that blocks apoptosis (PubMed:27806260).
Cellular Location	Cell membrane; Single-pass type II membrane protein. Cytoplasmic vesicle lumen Lysosome lumen. Note=Is internalized into multivesicular bodies of secretory lysosomes after phosphorylation by FGR and monoubiquitination (PubMed:17164290). Colocalizes with the SPPL2A protease at the cell membrane (PubMed:17557115) [FasL intracellular domain]: Nucleus. Note=The FasL ICD cytoplasmic form is translocated into the nucleus.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

REFERENCES

Alderson M.,et al.J. Exp. Med. 181:71-77(1995).
Takahashi T.,et al.Int. Immunol. 6:1567-1574(1994).
Schaetzlein C.E.,et al.Submitted (JUN-1995) to the EMBL/GenBank/DDBJ databases.
Mita E.,et al.Biochem. Biophys. Res. Commun. 204:468-474(1994).
Zeytun A.,et al.Submitted (JUL-2000) to the EMBL/GenBank/DDBJ databases.

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¥ 1,250.00

Cat# AP52211

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Fas Ligand Rabbit pAb