Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 488 Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker
  • Suitable for: Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm

Overview

  • Product name

    Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker
    See all Calreticulin primary antibodies

  • Description

    Alexa Fluor® 488 Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker

  • 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. This information is proprietary to Abcam and/or its suppliers.
    (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

    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.

    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 this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR3924

  • Isotype

    IgG

  • Research areas

Images

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

    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 (5 minutes), 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 1 hour. 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 labeled in blue with DAPI.

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

  • Flow Cytometry - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)
    Flow Cytometry - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158) Chaurasiya S et al Cancer Gene Ther. 2019 Jun 17. doi: 10.1038/s41417-019-0114-x. Fig 3a. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Cells were mock-infected or infected with TK-deleted CF33 virus at MOI 5 in a 6-well plate. The relatively high MOI (MOI 5) was used to ensure that all cells get infected by the virus. Eighteen hours post-infection, cells were detached using 10 mM EDTA and stained with AlexaFluor®488-conjugated anti-calreticulin antibody (ab196158; Abcam) or isotype antibody (ab199091; Abcam). Cells were fixed with 4% paraformaldehyde and analyzed on a flow cytometer (Accuri C6; BD).

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

    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 minutes) and then permeabilized with 0.1% PBS-Triton X-100 for 15 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab196158, 0.1 µg/ml dilution) for 30 minutes 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). Unlabeled 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 minutes), permeabilized with 0.1% PBS-Triton X-100 for 15 minutes under the same conditions.

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

    ab196158 staining Calreticulin in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed with 100% methanol (5 minutes), 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 1 hour. 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 labeled with DAPI (shown in blue).

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

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

    Overlay histogram showing HeLa cells (Human epithelial cell line from cervix adenocarcinoma) stained with ab196158 (red line).

    The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 minutes 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. Unlabeled 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 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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