Validated using a knockout cell line
Recombinant
RabMAb

Anti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 488) (ab196158)

Overview

  • Product name
    Anti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 488)
    See all Calreticulin primary antibodies
  • Description
    Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker (Alexa Fluor® 488)
  • Host species
    Rabbit
  • Conjugation
    Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Tested applications
    Suitable for: Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Monkey
  • Immunogen

    Synthetic peptide within Human Calreticulin aa 50-150. The exact sequence is proprietary.
    (Peptide available as ab180826)

  • Positive control
    • ICC/IF: HeLa cells, HAP1 cells (HAP1-CALR knockout cells used as negative cell line) Flow Cyt: HeLa cells, HAP1-WT cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com

    This product is a recombinant rabbit monoclonal antibody.

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. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer
    pH: 7.4
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Monoclonal
  • Clone number
    EPR3924
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab196158 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 1/50.

ab199091 - Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control with this antibody.

ICC/IF 1/1000.

This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min)

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.
  • Information by UniProt
  • Database links
  • 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

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-CALR knockout cells (red line) stained with ab196158. 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 (ab196158, 0.1µg/ml dilution) for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (ab199091) 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.

    This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • ab196158 staining Calreticulin (shown in green) 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 ab196158 at 1/500 dilution (shown in green) and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

  • ab196158 staining Calreticulin in HeLa cells. The cells were fixed with 100% methanol (5 min), 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 overnight at +4°C with ab196158 at 1/1000 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

  • Overlay histogram showing HeLa cells stained with ab196158 (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 (ab196158, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in HeLa fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • O'Leary MP  et al. Novel oncolytic chimeric orthopoxvirus causes regression of pancreatic cancer xenografts and exhibits abscopal effect at a single low dose. J Transl Med 16:110 (2018). Read more (PubMed: 29699566) »
  • Pan W  et al. A novel SMAC mimetic APG-1387 exhibits dual antitumor effect on HBV-positive hepatocellular carcinoma with high expression of cIAP2 by inducing apoptosis and enhancing innate anti-tumor immunity. Biochem Pharmacol 154:127-135 (2018). Human . Read more (PubMed: 29679556) »

See all 6 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Mouse Cell (GL261, Glioblastoma)
Permeabilization
No
Gating Strategy
Gated on only viable cells.
Specification
GL261, Glioblastoma
Fixation
No fixation was used.
Username

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Verified customer

Submitted Mar 10 2017

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