Anti-Chromogranin A antibody (ab85554)
Key features and details
- Rabbit polyclonal to Chromogranin A
- Suitable for: IHC-P, WB, IP, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Chromogranin A antibody
See all Chromogranin A primary antibodies -
Description
Rabbit polyclonal to Chromogranin A -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Horse, Cow, Dog, Pig, Monkey
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Immunogen
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Positive control
- Recombinant Human Chromogranin A protein (ab85486) can be used as a positive control in WB. This antibody gave a positive signal in human pancreas tissue lysate. This antibody gave a positive result when used in the following formaldehyde/methanol fixed cell line:PANC-1.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab85554 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 IHC-P Use a concentration of 5 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). IP Use a concentration of 5 µg/ml. ICC/IF Use a concentration of 1 - 5 µg/ml. Target
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Function
Pancreastatin strongly inhibits glucose induced insulin release from the pancreas. -
Sequence similarities
Belongs to the chromogranin/secretogranin protein family. -
Post-translational
modificationsSulfated on tyrosine residues and/or contains sulfated glycans.
O-glycosylated with core 1 or possibly core 8 glycans. -
Cellular localization
Secreted. Neuroendocrine and endocrine secretory granules. - Information by UniProt
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Database links
- Entrez Gene: 281070 Cow
- Entrez Gene: 100033828 Horse
- Entrez Gene: 1113 Human
- Entrez Gene: 12652 Mouse
- Entrez Gene: 397540 Pig
- Entrez Gene: 24258 Rat
- Omim: 118910 Human
- SwissProt: P05059 Cow
see all -
Form
According to the Swiss-Prot database, this protein has many different forms which correspond to various molecular weights; thus the possible variation from the predicted molecular weight of 51kDa. -
Alternative names
- beta Granin antibody
- betagranin (N-terminal fragment of chromogranin A) antibody
- catestatin antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (ab85554)IHC image of Chromogranin A staining in Human Pancreas Adenocarcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85554, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
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Immunoprecipitation - Anti-Chromogranin A antibody (ab85554)Chromogranin A was immunoprecipitated using 0.5mg Mouse Pancreas tissue lysate, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Pancreas tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85554.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 50kDa; Chromogranin A
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Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody (ab85554)ICC/IF image of ab85554 stained PANC-1 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 ab85554 at 5 µg/mL overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) used at a 1/250 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.
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Western blot - Anti-Chromogranin A antibody (ab85554)Anti-Chromogranin A antibody (ab85554) at 1 µg/ml + Human pancreas tissue lysate - total protein (ab29816) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDa
Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (ab85554)This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United KingdomIHC image of Chromogranin A staining in Monkey Pancreas sections, performed The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6). Sections were subjected to 2% H202O then Avidin-Biotin to block endogenous peroxidases/biotin. The section was then incubated with ab85554, 1/5000, for 2 hours at 12°C. The section was then counterstained with haematoxylin
Protocols
References (4)
ab85554 has been referenced in 4 publications.
- Dávalos-Salas M et al. Deletion of intestinal Hdac3 remodels the lipidome of enterocytes and protects mice from diet-induced obesity. Nat Commun 10:5291 (2019). PubMed: 31757939
- Moullé VS et al. Glucose and fatty acids synergistically and reversibly promote beta cell proliferation in rats. Diabetologia 60:879-888 (2017). PubMed: 28078385
- Hayase E et al. R-Spondin1 expands Paneth cells and prevents dysbiosis induced by graft-versus-host disease. J Exp Med 214:3507-3518 (2017). PubMed: 29066578
- Terry NA et al. Impaired enteroendocrine development in intestinal-specific Islet1 mouse mutants causes impaired glucose homeostasis. Am J Physiol Gastrointest Liver Physiol 307:G979-91 (2014). PubMed: 25214396
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (ab85554)IHC image of Chromogranin A staining in Human Pancreas Adenocarcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85554, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
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Immunoprecipitation - Anti-Chromogranin A antibody (ab85554)
Chromogranin A was immunoprecipitated using 0.5mg Mouse Pancreas tissue lysate, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Pancreas tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85554.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 50kDa; Chromogranin A
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Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody (ab85554)
ICC/IF image of ab85554 stained PANC-1 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 ab85554 at 5 µg/mL overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) used at a 1/250 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.
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Western blot - Anti-Chromogranin A antibody (ab85554)Anti-Chromogranin A antibody (ab85554) at 1 µg/ml + Human pancreas tissue lysate - total protein (ab29816) at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 50 kDa
Additional bands at: 72 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 20 minutes
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (ab85554) This image is courtesy of an abreview submitted by Carl Hobbs, King's College London, United Kingdom
IHC image of Chromogranin A staining in Monkey Pancreas sections, performed The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6). Sections were subjected to 2% H202O then Avidin-Biotin to block endogenous peroxidases/biotin. The section was then incubated with ab85554, 1/5000, for 2 hours at 12°C. The section was then counterstained with haematoxylin
