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    calreticulin-antibody-epr3924-bsa-and-azide-free-ab271865.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

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Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
  • Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3924] to Calreticulin - BSA and Azide free
  • Suitable for: IHC-P, Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human, African green monkey

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-Calreticulin antibody [EPR3924] - BSA and Azide free
    See all Calreticulin primary antibodies
  • Description

    Rabbit monoclonal [EPR3924] to Calreticulin - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human, African green monkey
    Predicted to work with: Rat, Monkey
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: HAP1 cells (HAP1-CALR as negative cell line) IHC-P: Human colon, kidney, liver, placenta, stomach, breast carcinoma and Papillary carcinoma of thyroid gland tissues; Mouse liver and Rat lung tissues.
  • General notes

    ab271865 is the carrier-free version of ab92516.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3924
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • ER

Associated products

  • Alternative Versions

    • Alexa Fluor® 555 Anti-Calreticulin antibody [EPR3924] (ab275117)
    • Alexa Fluor® 594 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab275343)
    • Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab271865 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 at an assay dependent concentration.

The use of a HRP/AP polymerized secondary antibody is recommended for enhanced staining.

 

Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
ICC/IF
Use at an assay dependent concentration.
Notes
IHC-P
Use at an assay dependent concentration.

The use of a HRP/AP polymerized secondary antibody is recommended for enhanced staining.

 

Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  

WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
ICC/IF
Use at an assay dependent concentration.

Target

  • Function

    Molecular calcium-binding chaperone promoting folding, oligomeric assembly and quality control in the ER via the calreticulin/calnexin cycle. This lectin interacts transiently with almost all of the monoglucosylated glycoproteins that are synthesized in the ER. Interacts with the DNA-binding domain of NR3C1 and mediates its nuclear export.
  • Sequence similarities

    Belongs to the calreticulin family.
  • Domain

    Can be divided into a N-terminal globular domain, a proline-rich P-domain forming an elongated arm-like structure and a C-terminal acidic domain. The P-domain binds one molecule of calcium with high affinity, whereas the acidic C-domain binds multiple calcium ions with low affinity.
    The interaction with glycans occurs through a binding site in the globular lectin domain.
    The zinc binding sites are localized to the N-domain.
    Associates with PDIA3 through the tip of the extended arm formed by the P-domain.
  • Cellular localization

    Endoplasmic reticulum lumen. Cytoplasm > cytosol. Secreted > extracellular space > extracellular matrix. Cell surface. Also found in cell surface (T cells), cytosol and extracellular matrix. Associated with the lytic granules in the cytolytic T-lymphocytes.
  • Target information above from: UniProt accession P27797 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 811 Human
    • Entrez Gene: 12317 Mouse
    • Entrez Gene: 64202 Rat
    • Omim: 109091 Human
    • SwissProt: P27797 Human
    • SwissProt: P14211 Mouse
    • SwissProt: P18418 Rat
    • Unigene: 515162 Human
    • Unigene: 1971 Mouse
    • Unigene: 467043 Mouse
    • Unigene: 974 Rat
    see all
  • Alternative names

    • Autoantigen RO antibody
    • CALR antibody
    • CALR protein antibody
    • CALR_HUMAN antibody
    • Calregulin antibody
    • Calreticulin antibody
    • cC1qR antibody
    • CRP55 antibody
    • CRT antibody
    • CRTC antibody
    • Endoplasmic reticulum resident protein 60 antibody
    • Epididymis secretory sperm binding protein Li 99n antibody
    • ERp60 antibody
    • FLJ26680 antibody
    • grp60 antibody
    • HACBP antibody
    • HEL S 99n antibody
    • RO antibody
    • Sicca syndrome antigen A (autoantigen Ro; calreticulin) antibody
    • Sicca syndrome antigen A antibody
    • SSA antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    ab92516 staining Calreticulin in wild-type HAP1 cells (top panel) and CALR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92516 at 1/500 and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    Alexa Fluor® 488 (ab196158) and Alexa Fluor® 647 (ab196159) conjugated versions are available for this clone.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Calreticulin  with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat lung.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-CALR knockout cells (red line) stained with ab92516. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab92516, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CALR knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. 

    Alexa Fluor® 488 (ab196158) and Alexa Fluor® 647 (ab196159) conjugated versions are available for this clone. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Calreticulin  with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on mouse liver.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining human breast carcinoma.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Formalin-fixed, paraffin-embedded normal human colon tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

  • Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) labelling Calreticulin with purified ab92516 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Overlay histogram showing HeLa cells stained with ab92516 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92516, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    ab92516, at 1/250 dilution, staining Calreticulin in paraffin embedded Human kidney tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    ab92516 showing negative staining in Normal human heart tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Formalin-fixed, paraffin-embedded normal human liver tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Formalin-fixed, paraffin-embedded normal human placenta tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Formalin-fixed, paraffin-embedded normal human stomach tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

    Formalin-fixed, paraffin-embedded Papillary carcinoma of human thyroid gland tissue.stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92516).

  • Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
    Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)

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

  • Datasheet download

    Download

Certificate of Compliance

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

References (0)

Publishing research using ab271865? Please let us know so that we can cite the reference in this datasheet.

ab271865 has not yet been referenced specifically in any publications.

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