CHGA Rabbit pAb

Blank control（black line）:SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-CHGA antibody (AP94805)
Dilution:2ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min 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.
Sample:
Lane 1: Mouse Adrenal gland tissue lysates
Lane 2: Rat Adrenal gland tissue lysates
Lane 3: Recombinant mouse CHGA protein, C-His (HEK293)(AP94805)
Lane 4: Mouse NIH/3T3 cell lysates（negative control）
Primary: Anti-CHGA (AP94805) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 33,76 kDa
Paraformaldehyde-fixed, paraffin embedded Mouse Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CHGA Polyclonal Antibody, Unconjugated (AP94805) at 1:600 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple, AP94805-Cy5), DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (Human duodenum); 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; Incubation with (CHGA) Polyclonal Antibody, Unconjugated (AP94805) at 1:600 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

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实验流程
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	P26339
Reactivity	Rat, Human, Mouse
Host	Rabbit
Clonality	Polyclonal
Calculated MW	51789 Da
Physical State	Liquid
Immunogen	Recombinant mouse CHGA protein
Epitope Specificity	272-463/463
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	Cytoplasmic vesicle, secretory vesicle lumen. Cytoplasmic vesicle, secretory vesicle membrane. Secreted. Note=Associated with the secretory granule membrane through direct interaction to SCG3 that in turn binds to cholesterol-enriched lipid rafts in intragranular conditions.
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 member of the chromogranin/secretogranin family of neuroendocrine secretory proteins. It is found in secretory vesicles of neurons and endocrine cells. This gene product is a precursor to three biologically active peptides; vasostatin, pancreastatin, and parastatin. These peptides act as autocrine or paracrine negative modulators of the neuroendocrine system. Other peptides, including chromostatin, beta-granin, WE-14 and GE-25, are also derived from the full-length protein. However, biological activities for these molecules have not been shown. [provided by RefSeq, Jul 2008].

Additional Information
Gene ID	12652
Other Names	Chromogranin-A, CgA, Pancreastatin, Beta-granin, WE-14, Catestatin, GE-25, Serpinin-RRG, Serpinin, AL26, p-Glu serpinin precursor, Chga
Dilution	IHC-P=1:500-1000,WB=1:500-2000,Flow-Cyt=2ug/Test,IF=1:500-1000,IHC-F=1:500-1000
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	Chga
Function	[Pancreastatin]: Strongly inhibits glucose induced insulin release from the pancreas. [Serpinin]: Regulates granule biogenesis in endocrine cells by up-regulating the transcription of protease nexin 1 (SERPINE2) via a cAMP-PKA-SP1 pathway. This leads to inhibition of granule protein degradation in the Golgi complex which in turn promotes granule formation (PubMed:21436258). Pyroglutaminated (pGlu)-serpinin exerts an antiapoptotic effect on cells exposed to oxidative stress (PubMed:21537909).
Cellular Location	Cytoplasmic vesicle, secretory vesicle. Cytoplasmic vesicle, secretory vesicle, neuronal dense core vesicle {ECO:0000250\|UniProtKB:P10354}. Secreted. Note=Associated with the secretory granule membrane through direct interaction to SCG3 that in turn binds to cholesterol-enriched lipid rafts in intragranular conditions

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