Overview

  • 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, Monkey
    Predicted to work with: Rat, Horse, Cow, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human Chromogranin A aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab237601, ab93753)

  • 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.

Properties

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).Can be blocked with Human Chromogranin A peptide (ab237601).
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

  • Function

    Pancreastatin strongly inhibits glucose induced insulin release from the pancreas.
  • Sequence similarities

    Belongs to the chromogranin/secretogranin protein family.
  • Post-translational
    modifications

    Sulfated 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
  • Database links

  • 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
    • CgA antibody
    • CHG A antibody
    • Chga antibody
    • chromofungin antibody
    • Chromogranin A antibody
    • Chromogranin A parathyroid secretory protein 1 antibody
    • Chromogranin A precursor antibody
    • ChromograninA antibody
    • CMGA_HUMAN antibody
    • ER-37 antibody
    • Pancreastatin antibody
    • Parastatin antibody
    • Parathyroid secretory protein 1 antibody
    • Pituitary secretory protein I antibody
    • Secretory protein I antibody
    • SP I antibody
    • SP-I antibody
    • SP1 antibody
    • SPI antibody
    • vasostatin 2 antibody
    • Vasostatin antibody
    • Vasostatin I antibody
    • Vasostatin II antibody
    • vasostatin-2 antibody
    see all

Images

  • 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
  • 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

  • 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.

  • 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
  • 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

    See Abreview

References

This product has been referenced in:

  • Moullé VS  et al. Glucose and fatty acids synergistically and reversibly promote beta cell proliferation in rats. Diabetologia 60:879-888 (2017). Read more (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). Read more (PubMed: 29066578) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Pancreas
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Jul 13 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Pancreas)
Specification
Pancreas
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Sep 01 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Adult Colon)
Specification
Adult Colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Sep 01 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Marmoset pancreas)
Specification
Marmoset pancreas
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Aug 26 2010

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