PE Anti-Calreticulin antibody [FMC 75] (ab83220)

Reviews (1)Q&A (5)References (6)
Flow Cytometry - PE Anti-Calreticulin antibody [FMC 75] (ab83220)

    Key features and details

    • PE Mouse monoclonal [FMC 75] to Calreticulin
    • Suitable for: Flow Cyt
    • Reacts with: Human, Monkey
    • Conjugation: PE. Ex: 488nm, Em: 575nm
    • Isotype: IgG1

    Overview

    • Product name

      PE Anti-Calreticulin antibody [FMC 75]
      See all Calreticulin primary antibodies

    • Description

      PE Mouse monoclonal [FMC 75] to Calreticulin

    • Host species

      Mouse

    • Conjugation

      PE. Ex: 488nm, Em: 575nm

    • Tested applications

      Suitable for: Flow Cytmore details

    • Species reactivity

      Reacts with: Human, Monkey

    • Immunogen

      Recombinant full length protein corresponding to Calreticulin.

    • Positive control

      • Jurkat cells.

    • General notes

      This product was changed from ascites to tissue culture supernatant on 31st May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

    Properties

    • Form

      Liquid

    • Storage instructions

      Shipped at 4°C. Store at +4°C.

    • Storage buffer

      pH: 7.20
      Preservative: 0.09% Sodium azide
      Constituent: PBS
    • Concentration information loading...

    • Purity

      Tissue culture supernatant

    • Clonality

      Monoclonal

    • Clone number

      FMC 75

    • Isotype

      IgG1

    • Light chain type

      kappa

    • Research areas

    Applications

    Our Abpromise guarantee covers the use of ab83220 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 Use at an assay dependent concentration.

    ab91357 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

     

    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

      • Flow Cytometry - PE Anti-Calreticulin antibody [FMC 75] (ab83220)
        Flow Cytometry - PE Anti-Calreticulin antibody [FMC 75] (ab83220)

        Intracellular staining of 106 Jurkat cells using ab83220 at a concentration of 10µg/ml.

        This image was generated using the ascites version of the product.

      References (6)

      Publishing research using ab83220? Please let us know so that we can cite the reference in this datasheet.

      ab83220 has been referenced in 6 publications.

      • Huang Y  et al. Comparison of the effects of photon, proton and carbon-ion radiation on the ecto-calreticulin exposure in various tumor cell lines. Ann Transl Med 7:542 (2019). PubMed: 31807524
      • Vera CA  et al. The nerve growth factor alters calreticulin translocation from the endoplasmic reticulum to the cell surface and its signaling pathway in epithelial ovarian cancer cells. Int J Oncol 50:1261-1270 (2017). PubMed: 28260038
      • Thierry S  et al. The DNA Repair Inhibitor Dbait Is Specific for Malignant Hematologic Cells in Blood. Mol Cancer Ther 16:2817-2827 (2017). PubMed: 28947503
      • Draganov D  et al. Modulation of P2X4/P2X7/Pannexin-1 sensitivity to extracellular ATP via Ivermectin induces a non-apoptotic and inflammatory form of cancer cell death. Sci Rep 5:16222 (2015). PubMed: 26552848
      • Angelova AL  et al. Complementary induction of immunogenic cell death by oncolytic parvovirus H-1PV and gemcitabine in pancreatic cancer. J Virol 88:5263-76 (2014). Flow Cyt ; Human . PubMed: 24574398
      • Wei H  et al. Bovine Herpesvirus Type 1 (BHV-1) U(L)49.5 Luminal Domain Residues 30 to 32 Are Critical for MHC-I Down-Regulation in Virus-Infected Cells. PLoS One 6:e25742 (2011). Flow Cyt ; Cow . PubMed: 22046246

      Customer reviews and Q&As

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      1-6 of 6 Abreviews or Q&A

      Flow Cytometry abreview for Anti-Calreticulin antibody [FMC 75] (Phycoerythrin)

       Excellent
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Mouse Cell (B16 melanoma)
      Permeabilization
      Yes - BD Perm/Fix kit was used for intracellular staining
      Gating Strategy
      Greater than 90% of cells were calreticulin+ compared to PE Isotype control
      Specification
      B16 melanoma
      Preparation
      Cell harvesting/tissue preparation method: Cells were trypinised to obtain a single cell suspension.
      Sample buffer: Perm/Fix Buffer
      Fixation
      Paraformaldehyde
      Read More

      Felicia Cao

      Verified customer

      Submitted Jul 27 2015

      Question

      I have a question regarding your PE-conjugated anti-Calreticulin antibody [FMC 75] (ab83220). I intend to use this product for detection of calreticulin on the cell surface. Do you know whether this mAb recognizes an epitope of calreticulin that is located intracellularly, or will it also recognize surface (ecto) calreticulin? The control stain on your website shows intracellular staining of HeLa cells, but I am specifically trying to detect calreticulin on the surface.

      Read More
      Answer

      According to http://www.uniprot.org/uniprot/P27797 Calreticulin is localized to the ER, cytoplasm, secreted, and on the cell surface, so rather than being a multipass membrane protein it is translocated to different regions of the cell. Thus, uniprot does not list specific amino acid ranges of the protein that are intracellular vs. extracellular when it is localized on the cell surface, and since it is most prevalent in the ER and cytoplasm our anti-Calreticulin antibodies were validated in ICC and flow with fixed, permeabilized cells. Thus, I am not sure whether or not our anti-Calreticulin antibodies, including ab22683, will bind to Calreticulin on the cell surface because I do not know what portion of the protein is extracellular vs. intracellular when it is surface-localized.

      Read More

      Question

      Hello, I ordered ab83220 anti-CALR antibody and I tested it in different human leukemic cells lines and different mutants for CALR. As in some cases I don't have membrane detection of CALR I was wondering if you know which part of the protein is recognize by the antibody (C-term or N-term). Indeed, I would help to confirm my results and go further in biological characterization.

      Read More
      Answer

      The epitope that this calreticulin antibody recognizes has not been mapped but it is expected that the epitope is on an extracellular region. The unconjugated version of this clone, ab22863, was used for staining live cells for flow cytometry in PubMed: 18689689, http://www.pnas.org/content/105/33/11784.long.

      It is not clear how much of the protein is extracellular but it is expected to include the N terminus, based on studies of cell-surface CRT. The review FASEB J. 2010 Mar;24(3):665-83. at http://www.fasebj.org/content/24/3/665.long discusses these studies.

      Read More

      Question

      Is the immunogen a fragment or full length protein?

      Read More
      Answer

      The immunogen for ab83220 is the full length recombinant protein.

      Read More

      Question

      Conjugated abs gives signal, whereas the non-conjugated with secondary fails to do so. Secondary works fine with other abs.
      Are checking at the moment if pos. signal is non-specific with siRNA.

      Read More
      Answer

      Vielen Dank für Ihren Anruf.

      Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie am Telefon besprochen habe ich eine kostenlose Ersatzlieferung mit einer anderen Lot# in Auftrag gegeben. Sie hat die Referenznummer 123456 und sollte Montag bei Ihnen ankommen.

      Als Anhang finden Sie die beiden Fragebögen für die Flow -Analyse der beiden Antikörper.

      Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

      Read More

      Question

      Inquiry: Over a year ago I bought a direct mouse antibody to calreticulin conjugated with PE (ab83220) and used it to detect CRT externalization during the apoptosis of hamster's melanoma cells. This antibody worked well. This year I would like to order the next portion of this antibody but to my surprise in the description of the antibody not conjugated with PE (ab22683) I found an information that antibody of this clone doe not react with hamster's cells. My question is what hamster's cells have been used to make this conclusion ? I feel a little confused, what I detected then ? Sincerely yours

      Read More
      Answer

      Thank you for your enquiry regarding ab22683.

      We have removed Hamster from the "does not react" species list, this seems to be an error.

      We apologize for any inconvenience caused and thank you for bringing this matter to our attention.

      If you need any further assistance in the future, please do not hesitate to contact me.

      Read More

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