SPAG17 Rabbit pAb

Blank control: HL60.
Primary Antibody (green line): Rabbit Anti-SPAG17 antibody (AP94693)
Dilution: 1 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1 µg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Tissue/cell: mouse brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-SPAG17 Polyclonal Antibody, Unconjugated(AP94693) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control: HL60. Primary Antibody (green line): Rabbit Anti-SPAG17 antibody (AP94693) Dilution: 1 µg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PE Dilution: 1 µg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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背景知识

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	IHC-P, IHC-F, IF
Primary Accession	Q6Q759
Reactivity	Human, Mouse
Predicted	Rat
Host	Rabbit
Clonality	Polyclonal
Calculated MW	251742 Da
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from human SPAG17
Epitope Specificity	741-840/2223
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	Cytoplasm. Cytoplasm > cytoskeleton > flagellum axoneme. Detected in the cytoplasm of round spermatids and in condensing spermatids. Localized to the central pair of the sperm flagellar axoneme. Colocalizes with SPAG6 on microtubules.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	SPAG17 (sperm associated antigen 17), also known as PF6, is a 2,223 amino acid cytoplamic protein that colocalizes with SPAG6 to microtubles. Highly expressed in testis and in organs that contain cilia-bearing cells including brain, oviduct, lung, and uterus, SPAG17 may be important for the structural integrity of the central apparatus of the sperm axoneme. SPAG17 contains two LRR (leucine-rich) repeats and may also participate in flagellar motility and male fertility.

Additional Information
Gene ID	200162
Other Names	Sperm-associated antigen 17, Projection protein PF6 homolog, SPAG17
Dilution	IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/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	SPAG17
Function	Component of the central pair apparatus of ciliary axonemes. Plays a critical role in the function and structure of motile cilia. May play a role in endochondral bone formation, most likely because of a function in primary cilia of chondrocytes and osteoblasts (By similarity). Essential for normal spermatogenesis and male fertility (By similarity). Required for normal manchette structure, transport of proteins along the manchette microtubules and formation of the sperm head and flagellum (By similarity). Essential for sperm flagellum development and proper assembly of the respiratory motile cilia central pair apparatus, but not the brain ependymal cilia (By similarity).
Cellular Location	Cytoplasm {ECO:0000250\|UniProtKB:Q5S003}. Cytoplasm, cytoskeleton, flagellum axoneme {ECO:0000250\|UniProtKB:Q5S003}. Cytoplasmic vesicle, secretory vesicle, acrosome {ECO:0000250\|UniProtKB:Q5S003}. Golgi apparatus {ECO:0000250\|UniProtKB:Q5S003}. Cytoplasm, cytoskeleton {ECO:0000250\|UniProtKB:Q5S003}. Note=Detected in the cytoplasm of round spermatids and in condensing spermatids. Localized to the central pair of the sperm flagellar axoneme. Colocalizes with SPAG6 on microtubules (By similarity). Localizes to the manchette in elongating spermatids (By similarity). {ECO:0000250\|UniProtKB:Q5S003}
Tissue Location	Highly expressed in testis. Expressed in organs that contain cilia-bearing cells including brain, oviduct, lung, and uterus.

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