Recombinant Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Alexa Fluor® 647 Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker
- Suitable for: ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
- Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
Related conjugates and formulations
Overview
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Product name
Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker
See all Calreticulin primary antibodies -
Description
Alexa Fluor® 647 Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker -
Host species
Rabbit -
Conjugation
Alexa Fluor® 647. Ex: 652nm, Em: 668nm -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab180826) -
Positive control
- ICC/IF: HeLa and HAP1 cells (HAP1-CALR knockout cells used as a negative cell line). Flow Cyt (intra): HeLa and HAP1-WT cells.
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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
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.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3924 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- HRP Anti-Calreticulin antibody [EPR3924] - ER Marker (ab195511)
- Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)
- PE Anti-Calreticulin antibody [EPR3924] - ER Marker (ab209577)
- Alexa Fluor® 405 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab210431)
- Anti-Calreticulin antibody [EPR3924] - Low endotoxin, Azide free (ab211962)
- 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)
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab196159 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 |
---|---|---|
ICC/IF | (2) |
1/50 - 1/500.
|
Flow Cyt (Intra) |
1/50.
|
Notes |
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ICC/IF
1/50 - 1/500. |
Flow Cyt (Intra)
1/50. |
Target
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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
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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
see all -
Alternative names
- Autoantigen RO antibody
- CALR antibody
- CALR protein antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)
ab196159 staining Calreticulin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabiliszd in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab196159 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)
ab196159 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 ab196159 at 1/500 dilution (shown in red) and ab195887at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)
Overlay histogram showing HAP1 wildtype (green line) and HAP1-CALR knockout cells (red line) stained with ab196159. 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 (ab196159, 0.1µg/ml dilution) for 30 min at 22°C.
A rabbit monoclonal IgG isotype control antibody (ab199093) 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 40 mW Red laser (640nm) and 670/14 bandpass filter. -
Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)Overlay histogram showing HeLa cells stained with ab196159 (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 (ab196159, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG [EPR25A] Alexa Fluor® 647 (ab199093) 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 solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (22)
ab196159 has been referenced in 22 publications.
- von Hofsten S et al. The marine natural product mimic MPM-1 is cytolytic and induces DAMP release from human cancer cell lines. Sci Rep 12:15586 (2022). PubMed: 36114339
- Yu X et al. Novel formulation of c-di-GMP with cytidinyl/cationic lipid reverses T cell exhaustion and activates stronger anti-tumor immunity. Theranostics 12:6723-6739 (2022). PubMed: 36185614
- Fujimoto T et al. Cathepsin B is highly expressed in pancreatic cancer stem-like cells and is associated with patients' surgical outcomes. Oncol Lett 21:30 (2021). PubMed: 33240436
- Wang Y et al. Copper-Based Nanoscale Coordination Polymers Augmented Tumor Radioimmunotherapy for Immunogenic Cell Death Induction and T-Cell Infiltration. Small N/A:e2006231 (2021). PubMed: 33522120
- He ZD et al. Anti-PD-L1 mediating tumor-targeted codelivery of liposomal irinotecan/JQ1 for chemo-immunotherapy. Acta Pharmacol Sin 42:1516-1523 (2021). PubMed: 33311600