Name: Anti-Chromogranin A antibody [RM1025]
Brand: RabMAb
SKU: ab283265
Rating: 5 (1 reviews)

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Western blot - Anti-Chromogranin A antibody [RM1025] (ab283265)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit recombinant multiclonal [RM1025] to Chromogranin A
Suitable for: WB, ICC/IF, Flow Cyt (Intra), IHC-P, IHC-Fr, ELISA, IP
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-Chromogranin A antibody [RM1025]
See all Chromogranin A primary antibodies
Description

Rabbit recombinant multiclonal [RM1025] to Chromogranin A
Host species

Rabbit
Tested applications

Suitable for: WB, ICC/IF, Flow Cyt (Intra), IHC-P, IHC-Fr, ELISA, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

This product was produced with the following immunogens:
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human adrenal gland, SH-SY5Y, Neuro-2a, Mouse adrenal gland, Rat adrenal gland, PC-12 lysates. IHC-P: Human pancreas, Human stomach, Mouse pancreas, Mouse adrenal gland, Rat pancreas, Rat adrenal gland tissues. IHC-Fr: Mouse colon, Rat colon tissues. ICC/IF: SH-SY5Y, Neuro-2a cells. Flow Cyt (Intra): SH-SY5Y, Neuro-2a cells. IP: SH-SY5Y, Neuro-2a cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Clonality

Recombinant Multiclonal
Clone number

RM1025
Isotype

IgG
Research areas
- Neuroscience
- Processes

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Isotype control
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283265 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
WB		1/1000. Predicted molecular weight: 50 kDa.
ICC/IF		1/50.
Flow Cyt (Intra)		1/500.
IHC-P	(1)	1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr		1/500.
ELISA		Use a concentration of 0.125 µg/ml.
IP		1/30.

Notes
WB 1/1000. Predicted molecular weight: 50 kDa.
ICC/IF 1/50.
Flow Cyt (Intra) 1/500.
IHC-P 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr 1/500.
ELISA Use a concentration of 0.125 µg/ml.
IP 1/30.

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.
Target information above from: UniProt accession P10645 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 1113 Human
- Entrez Gene: 12652 Mouse
- Entrez Gene: 24258 Rat
- Omim: 118910 Human
- SwissProt: P10645 Human
- SwissProt: P26339 Mouse
- SwissProt: P10354 Rat
- Unigene: 150793 Human
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

Western blot - Anti-Chromogranin A antibody [RM1025] (ab283265)

All lanes : Anti-Chromogranin A antibody [RM1025] (ab283265) at 1/1000 dilution

Lane 1 : Human adrenal gland lysate
Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 4 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 50 kDa
Observed band size: 20,75 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Negative control: Jurkat.

The molecular weight observed is consistent with what has been described in the literature (PMID: 14734658, 21052719).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time: 3 min
Western blot - Anti-Chromogranin A antibody [RM1025] (ab283265)

All lanes : Anti-Chromogranin A antibody [RM1025] (ab283265) at 1/1000 dilution

Lane 1 : Mouse adrenal gland lysate
Lane 2 : Rat adrenal gland lysate
Lane 3 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 50 kDa
Observed band size: 20,75 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Negative control: NIH/3T3. (PMID: 7615829)

Exposure time: Lane 1-2: 5 seconds Lane 3-5: 92 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in human pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the endocrine cells in human stomach. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in mouse pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded mouse adrenal gland tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining in the mouse adrenal gland. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on the islet in rat pancreas. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of paraffin-embedded rat adrenal gland tissue labelling Chromogranin A with ab283265 at 1/5000 dilution (0.107 µg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining in the rat adrenal gland. The section was incubated with ab283265 overnight at 4℃. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0)
Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse colon (fresh) tissue labeling Chromogranin A with ab283265 at 1/500 dilution (1.074 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/ml) (Green). Positive satining on mouse colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.
Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat colon (fresh) tissue labeling Chromogranin A with ab283265 at 1/500 dilution (1.074 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (2 µg/ml) (Green). Positive staining on rat colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.
Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labelling Chromogranin A with ab283265 at 1/50 dilution (10.74 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing cytoplasmic staining in subsets of SH-SY5Y cells.

Negative control: Jurkat.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed.
Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [RM1025] (ab283265)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labelling Chromogranin A with ab283265 at 1/50 dilution (10.74 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing cytoplasmic staining in of Neuro-2a cells.

Negative control: NIH/3T3 (PMID: 7615829).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/ml) (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed.
Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte / Left) and SH-SY5Y (Human neuroblastoma epithelial cell / Right) cells labelling Chromogranin A with ab283265 at 1/500 dilution (0.1 µg) compared with an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control: Jurkat.
Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [RM1025] (ab283265)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast / Left) and Neuro-2a (Mouse neuroblastoma neuroblast / Right) cells labelling Chromogranin A with ab283265 at 1/500 dilution (0.1 µg / Red) compared with a Rabbit monoclonal IgG (ab172730 / Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control: NIH/3T3. (PMID: 7615829)
Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (ab283265)

Chromogranin A was immunoprecipitated from 0.35 mg SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate with ab283265 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283265 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug

Lane 2: ab283265 IP in SH-SY5Y whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283265 in SH-SY5Y whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 30 seconds

Lower bands could be cleavage fragment. (PMID: 17991725)
Immunoprecipitation - Anti-Chromogranin A antibody [RM1025] (ab283265)

Chromogranin A was immunoprecipitated from 0.35 mg Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate with ab283265 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283265 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate 10ug

Lane 2: ab283265 IP in Neuro-2a whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283265 in Neuro-2a whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 min

Lower bands could be cleavage fragment. (PMID: 17991725)
ELISA - Anti-Chromogranin A antibody [RM1025] (ab283265)

ELISA using ab283265 at varying antibody concentrations and antigen concentration at 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Anti-Chromogranin A antibody [RM1025] (ab283265)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab283265? Please let us know so that we can cite the reference in this datasheet.

ab283265 has been referenced in 1 publication.

Song H et al. Single-cell analysis of hepatoblastoma identifies tumor signatures that predict chemotherapy susceptibility using patient-specific tumor spheroids. Nat Commun 13:4878 (2022). PubMed: 36008377

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Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Gut & Pancreas)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6

Permeabilization

Specification

Gut & Pancreas

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 20°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Mary Green

Verified customer

Submitted Oct 31 2022

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Form

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (1)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Chromogranin A antibody [RM1025]

Post-translational
modifications