Anti-Secretogranin V antibody (ab22699)
- Datasheet
- References (3)
- Protocols
Overview
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Product name
Anti-Secretogranin V antibody
See all Secretogranin V primary antibodies -
Description
Rabbit polyclonal to Secretogranin V -
Host species
Rabbit -
Tested applications
Suitable for: IP, ICC/IF, WB, IHC-P, IHC-Frmore details -
Species reactivity
Reacts with: Rat, Human, Pig
Predicted to work with: Mouse, Chicken, Xenopus laevis, Zebrafish -
Immunogen
Synthetic peptide:
EQLGIARPRV EYPAHQA
conjugated to BSA by a CDI linker, corresponding to N terminal amino acids 23-39 of Pig Secretogranin V (45-61 of pro-Pig Secretogranin V).
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.065% Sodium azide -
Concentration information loading...
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Purification notes
Partially purified, caprylic acid and ammonium sulphate precipitative clean-up. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab22699 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
IP | Use at an assay dependent concentration. | |
ICC/IF | Use a concentration of 5 µg/ml. | |
WB | Use at an assay dependent concentration. Detects a band of approximately 25 kDa. | |
IHC-P | Use at an assay dependent concentration. | |
IHC-Fr | Use at an assay dependent concentration. PubMed: 20434189 |
Target
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Relevance
Known variously as APPG, 7B2 and neuroendocrine protein I, Secretogranin V was originally isolated from porcine pituitary in 1983. It soon became evident that Secretogranin V-like immunoreactants were present in many cell types from a variety of species, and it is now recognised to be widely distributed in neural and endocrine tissues. In the early 1990s full length (27kDa) Secretogranin V was shown to be a potent inhibitor of prohormone convertase PC2 activity, a subtilisin-like enzyme responsible for the proteolytic cleavage of many neuroendocrine precursor molecules, including proenkephalin, POMC and gastrin-17. mRNA distribution analysis revealed that in adult rat brain, expression of Secretogranin V mRNA was pan-neuronal, whereas PC2 mRNA was exclusively neuronal, but more restricted. PC2+/Secretogranin V- cells were not encountered in the adult, but were common in developing rat brain. These findings suggest that Secretogranin V may have intracellular functions other than PC2 maturation in certain cells, although some recent work shows a discordancy between mRNA expression and enzymic activity. The mechanism for PC2 maturation appears to involve pro-Secretogranin V binding to inactive pro-PC2 via its polyproline motif (116 PPNPCP 121) in the endoplasmic reticulum and chaperoning pro-PC2 to a later secretory pathway compartment where maturation of PC2 proceeds once pro-Secretogranin V is proteolytically processed by a furin-like convertase into a 21kDa fragment and a C-terminal peptide. It is the C-terminal peptide that potently inhibits maturation of pro-PC2 until the complex is subjected to a decreasing pH gradient along the secretory pathway. It seems likely that mature PC2 then cleaves the CT peptide at its internal lys-lys site. Secretogranin V knockout mice develop a severe Cushings-like phenotype and exhibit multiple metabolic and hormonal abnormalities, indicating that Secretogranin V is required for activation of PC2 in vivo. Conversely, PC2-null mice appear to be viable. -
Cellular localization
Neuroendocrine and endocrine secretory granules. -
Database links
- Entrez Gene: 6447 Human
- Entrez Gene: 20394 Mouse
- Entrez Gene: 397110 Pig
- Entrez Gene: 25719 Rat
- Omim: 173120 Human
- SwissProt: P05408 Human
- SwissProt: P12961 Mouse
- SwissProt: P01165 Pig
see all -
Alternative names
- 7B2 antibody
- 7B2 protein antibody
- APPG antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Secretogranin V antibody (ab22699)ab75197 (1µg/ml) staining secretogranin V in human pancreas using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of Islets of Langerhans.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
ICC/IF image of ab22699 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22699, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Zheng B et al. A three-gene panel that distinguishes benign from malignant thyroid nodules. Int J Cancer 136:1646-54 (2015). Read more (PubMed: 25175491) »
- Sato A et al. Distribution of 7B2 (secretogranin V)-like immunoreactivity in the Japanese red-bellied newt (Cynops pyrrhogaster) pituitary. Tissue Cell 42:176-80 (2010). IHC-Fr ; Newt . Read more (PubMed: 20434189) »