Recombinant Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22537-248] to Chromogranin A
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Fr, ICC/IF
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-Chromogranin A antibody [EPR22537-248]
See all Chromogranin A primary antibodies -
Description
Rabbit monoclonal [EPR22537-248] to Chromogranin A -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Fr, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Neuro-2a and PC12 whole cell lysates; mouse serum; mouse adrenal gland lysate; rat adrenal gland lysate. IHC-P: Mouse pancreas tissue; rat pancreas tissue. IHC-Fr: Mouse colon tissue; rat colon tissue. ICC/IF: Neuro-2a and PC12 cells. Flow Cyt (intra): Neuro-2a cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22537-248 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab254322 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 |
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Flow Cyt (Intra) |
1/60.
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WB |
1/1000. Detects a band of approximately 20, 75 kDa (predicted molecular weight: 50 kDa).
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr |
1/500.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
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ICC/IF |
1/100.
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Notes |
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Flow Cyt (Intra)
1/60. |
WB
1/1000. Detects a band of approximately 20, 75 kDa (predicted molecular weight: 50 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
1/500. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
ICC/IF
1/100. |
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: 12652 Mouse
- Entrez Gene: 24258 Rat
- SwissProt: P26339 Mouse
- SwissProt: P10354 Rat
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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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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Chromogranin A with ab254322 at 1/100 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in Neuro-2a cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
Immunohistochemical analysis of paraffin-embedded rat panceas tissue labeling Chromogranin A with ab254322 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat pancreas islet (PMID: 22412953) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab254322 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-Chromogranin A antibody [EPR22537-248] (ab254322) at 1/1000 dilution
Lane 1 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 3 : Mouse serum
Lane 4 : Mouse adrenal gland lysate
Lane 5 : Rat adrenal gland lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 50 kDa
Observed band size: 20,75 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 15 seconds; Lane 2: 3.25 seconds; Lane 3:26 seconds; Lane 4: 3.25 seconds; Lane 5: 15 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 14734658, 21052719).
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Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Chromogranin A with ab254322 at 1/60 (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Chromogranin A with ab254322 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse pancreas islet (PMID: 22412953) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab254322 for 30 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat colon tissue labeling Chromogranin A with ab254322 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/1000 dilution (green). Positive staining on enteroendocrine cells of mouse colon (PMID: 27287799) is observed. The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell line) cells labeling Chromogranin A with ab254322 at 1/100 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in PC-12 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse colon tissue labeling Chromogranin A with ab254322 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/1000 dilution (green). Positive staining on enteroendocrine cells of mouse colon (PMID: 27287799). is observed. The nuclear counter stain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab254322 has not yet been referenced specifically in any publications.