GPR49 Rabbit pAb

Blank control (blue line):Hela(blue).
Primary Antibody (green line): Rabbit Anti-GPR49 antibody(AP94473),
Dilution: 3 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 µg /test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Sample:
Brain (Mouse) Lysate at 40 ug
Primary: Anti-GPR49 (AP94473) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 98 kD
Observed band size: 98 kD
Tissue/cell: mouse colon carcinoma; 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-GPR49/LGR5 Polyclonal Antibody, Unconjugated(AP94473) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

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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	WB, IHC-P, IHC-F, IF
Primary Accession	Q9Z1P4
Reactivity	Human, Mouse
Predicted	Rat
Host	Rabbit
Clonality	Polyclonal
Calculated MW	99666 Da
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from mouse GPR49
Epitope Specificity	810-907/907
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; Multi-pass membrane protein.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	The protein encoded by this gene is a leucine-rich repeat-containing receptor (LGR) and member of the G protein-coupled, 7-transmembrane receptor (GPCR) superfamily. The encoded protein is a receptor for R-spondins and is involved in the canonical Wnt signaling pathway. This protein plays a role in the formation and maintenance of adult intestinal stem cells during postembryonic development. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Sep 2015]

Additional Information
Gene ID	14160
Other Names	Leucine-rich repeat-containing G-protein coupled receptor 5, G-protein coupled receptor 49, Orphan G-protein coupled receptor FEX, Lgr5, Fex {ECO:0000303\|PubMed:9920770}, Gpr49
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	Lgr5
Synonyms	Fex {ECO:0000303\|PubMed:9920770}, Gpr49
Function	Receptor for R-spondins that potentiates the canonical Wnt signaling pathway and acts as a stem cell marker of the intestinal epithelium and the hair follicle. Upon binding to R-spondins (RSPO1, RSPO2, RSPO3 or RSPO4), associates with phosphorylated LRP6 and frizzled receptors that are activated by extracellular Wnt receptors, triggering the canonical Wnt signaling pathway to increase expression of target genes. In contrast to classical G protein-coupled receptors, does not activate heterotrimeric G proteins to transduce the signal. Involved in the development and/or maintenance of the adult intestinal stem cells during postembryonic development.
Cellular Location	Cell membrane; Multi-pass membrane protein. Golgi apparatus, trans-Golgi network membrane; Multi-pass membrane protein. Note=Rapidly and constitutively internalized to the trans-Golgi network at steady state.
Tissue Location	Expressed in the intestinal epithelium (at protein level) (PubMed:22510880). Expressed in the gonads, the adrenal gland, and in the brain. In the central nervous system expression is restricted to the olfactory bulb. In the adrenal gland detected only in the neural-crest derived chromaffin cells of the medulla, but not in the cells of the adrenal cortex. In the gonads, the expression is high in Graafian follicle, but absent from primary and secondary follicles In the intestine, exclusively expressed in cycling crypt base columnar cells. Expressed in the lower bulge and secondary germ area of telogen hair follicles and in the lower outer root sheath of anagen hair follicle.

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