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    chromogranin-a-antibody-epr22537-248-ab254322.pdf

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RecombinantRabMAb

Recombinant Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

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Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Western blot - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
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Anti-SLC7A7 antibody (ab236669)
Protein
Product image
Recombinant Human Chromogranin A protein (ab85486)

View more associated products

Overview

  • 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.

  • 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.
  • 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.

Properties

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22537-248
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Tumor Associated
    • Signal Transduction
    • Growth Factors/Hormones
    • Insulin / Insulin-like
    • Cancer
    • Growth factors
    • Insulin and insulin-like
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Chromogranin A antibody [EPR22537-248] - BSA and Azide free (ab255284)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunohistochemistry kits

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Chromogranin A protein (ab85486)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

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
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.
Notes
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.
Application notes
Is unsuitable for IP.

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.
  • 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: 12652 Mouse
    • Entrez Gene: 24258 Rat
    • SwissProt: P26339 Mouse
    • SwissProt: P10354 Rat
    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      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.

       

    • Western blot - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Western blot - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      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).

    • Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Flow Cytometry (Intracellular) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

      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. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      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.

       

    • Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Immunocytochemistry/ Immunofluorescence - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

      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.

    • Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

      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).

    • Anti-Chromogranin A antibody [EPR22537-248] (ab254322)
      Anti-Chromogranin A antibody [EPR22537-248] (ab254322)

    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.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab254322? Please let us know so that we can cite the reference in this datasheet.

    ab254322 has not yet been referenced specifically in any publications.

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