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  1. Link

    products/primary-antibodies/calreticulin-antibody-fmc-75-ab22683.pdf

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Anti-Calreticulin antibody [FMC 75] (ab22683)

  • Datasheet
  • SDS
Reviews (22)Q&A (16)References (71)

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Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [FMC 75] (ab22683)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [FMC 75] (ab22683)
  • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [FMC 75] (ab22683)

Key features and details

  • Mouse monoclonal [FMC 75] to Calreticulin
  • Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

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

    Mouse monoclonal [FMC 75] to Calreticulin
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Chinese hamster
  • Immunogen

    Fusion protein corresponding to Calreticulin. Calreticulin-maltose binding fusion protein.
    Database link: P27797

  • Positive control

    • WB: HeLa whole cell lysate. IHC-P: Human placenta tissue. ICC/IF: HeLa cell lysate. Flow: HeLa cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 22nd 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

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

    Protein G purified
  • Purification notes

    Purified from TCS.
  • Clonality

    Monoclonal
  • Clone number

    FMC 75
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • ER
    • Signal Transduction
    • Protein Trafficking
    • Organelle Proteins

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Related Products

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab22683 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)
Use at an assay dependent concentration.

(Additional resource: PMID - 18689689)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF (10)
Use at an assay dependent concentration.

Fix and permeabilize cells prior to staining.

IHC-P (2)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

(Additional resource: PMID - 18689689)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF
Use at an assay dependent concentration.

Fix and permeabilize cells prior to staining.

IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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
    • Omim: 109091 Human
    • SwissProt: Q8K3H7 Chinese hamster
    • SwissProt: P27797 Human
    • Unigene: 515162 Human
    • 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 [FMC 75] (ab22683)
      Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [FMC 75] (ab22683)

      ICC/IF image of ab22683 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22683, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [FMC 75] (ab22683)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [FMC 75] (ab22683)

      Ab22683 staining human placenta. Staining is localised to the cytoplasm.
      Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
      Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

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

    • Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [FMC 75] (ab22683)
      Flow Cytometry (Intracellular) - Anti-Calreticulin antibody [FMC 75] (ab22683)

      Overlay histogram showing HeLa cells stained with ab22683 (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 (ab22683, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

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

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (71)

    Publishing research using ab22683? Please let us know so that we can cite the reference in this datasheet.

    ab22683 has been referenced in 71 publications.

    • Blomme A  et al. THEM6-mediated reprogramming of lipid metabolism supports treatment resistance in prostate cancer. EMBO Mol Med 14:e14764 (2022). PubMed: 35014179
    • Saumell-Esnaola M  et al. Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization. Int J Mol Sci 23:N/A (2022). PubMed: 35270001
    • Zeng TM  et al. Immune-related RNA signature predicts outcome of PD-1 inhibitor-combined GEMCIS therapy in advanced intrahepatic cholangiocarcinoma. Front Immunol 13:943066 (2022). PubMed: 36159865
    • Cui H  et al. Re-expression of glucuronyl C5-epimerase in the mutant MEF cells increases heparan sulfate epimerization but has no influence on the Golgi localization and enzymatic activity of 2-O-sulfotransferase. Glycobiology N/A:N/A (2021). PubMed: 33755115
    • Ercan H  et al. Alterations of the Platelet Proteome in Lung Cancer: Accelerated F13A1 and ER Processing as New Actors in Hypercoagulability. Cancers (Basel) 13:N/A (2021). PubMed: 34066760
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 38 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Panc1 Tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA ph9
    Permeabilization
    No
    Specification
    Panc1 Tumor
    Blocking step
    Bloxall as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Dec 27 2021

    Western blot abreview for Anti-Calreticulin antibody [FMC 75]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293T)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    HEK293T
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Mar 10 2021

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HBE)
    Permeabilization
    Yes - 0.5% Triton in PBS
    Specification
    HBE
    Fixative
    Paraformaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Mar 05 2021

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Hamster Cell (CHO cells)
    Permeabilization
    Yes - Saponin 0.05% in PBS
    Specification
    CHO cells
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Sep 17 2019

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (OVCAR-3 ovary cancer cells)
    Permeabilization
    No
    Gating Strategy
    no gating
    Specification
    OVCAR-3 ovary cancer cells
    Preparation
    Cell harvesting/tissue preparation method: cell scrapped with PBS, and spun down
    Sample buffer: PBS with 0.5% BSA
    Fixation
    Formaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Mar 14 2019

    Immunocytochemistry abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (OV90 cells)
    Permeabilization
    Yes - 0.1% TritonX-100
    Specification
    OV90 cells
    Blocking step
    BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Jun 22 2018

    Western blot abreview for Anti-Calreticulin antibody [FMC 75]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Liver Extract)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin)
    Loading amount
    20 µg
    Specification
    Liver Extract
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Oct 25 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (B16 tumors taken out of C57bl mice)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6
    Permeabilization
    No
    Specification
    B16 tumors taken out of C57bl mice
    Blocking step
    Innovex Background Buster as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Sep 19 2017

    Western blot abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (OV90 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin gel)
    Loading amount
    800000 cells
    Specification
    OV90 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Aug 19 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Calreticulin antibody [FMC 75]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (B16 cell line)
    Permeabilization
    Yes - 0.25% Triton X-100
    Specification
    B16 cell line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    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.

    Abcam user community

    Verified customer

    Submitted Aug 12 2016

    1-10 of 38 Abreviews or Q&A

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