Product nameAnti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 647)
See all Calreticulin primary antibodies
DescriptionRabbit monoclonal [EPR3924] to Calreticulin - ER Marker (Alexa Fluor® 647)
ConjugationAlexa Fluor® 647. Ex: 652nm, Em: 668nm
Tested applicationsSuitable for: Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Calreticulin aa 50-150. The exact sequence is proprietary.
(Peptide available as
- ICC/IF: HeLa and HAP1 cells (HAP1-CALR knockout cells used as a negative cell line). Flow Cyt: HeLa and HAP1-WT cells.
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 email@example.com.
Storage instructionsStore at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
PurityImmunogen affinity purified
- Anti-Calreticulin antibody [EPR3924] - ER Marker (HRP) (ab195511)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 488) (ab196158)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (Phycoerythrin) (ab209577)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 405) (ab210431)
- Anti-Calreticulin antibody [EPR3924] - Low endotoxin, Azide free (ab211962)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
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.
|ICC/IF||1/50 - 1/500.|
FunctionMolecular 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 similaritiesBelongs to the calreticulin family.
DomainCan 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 localizationEndoplasmic 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
- Autoantigen RO antibody
- CALR antibody
- CALR protein antibody
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).
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.
Visualization of CRT expression was performed in KPC cells added to 8-well chamber slides (Lab-Tek®). Each well contained 1 × 104 KPC cells in 0.4 mL of culture medium. After incubation with 50 µM Cis, 50 µM OX, and 1 µM DOX for 4 h, cells were fixed and washed 3 times. Cells were stained with an Alexa Fluor® 647-conjugated anti-CRT antibody (ab196159, 1/500, Abcam) for 30 min, followed by co-staining with 5 μg/mL Alexa Fluor® 488-conjugated wheat germ agglutinin (WGA) to visualize the cell surface membrane. Slides were mounted with Hoechst 33,342 nuclear dye and visualized under a Leica SP8-SMD confocal microscope. High magnification images were obtained under the 63 × objective lens.
(Panel e) Flow cytometric analysis for the cell surface CRT in PMA-stimulated Jurkat cells.
Jurkat cells were stimulated with PMA (100 ng/mL) and dimethyl sulfoxide or compounds for 30 min. Cells were washed with PBS containing 0.3% FBS and fixed with 3.7% formaldehyde at 37 °C for 10 min. After washing with PBS (0.3% FBS), Alexa Fluor 647-conjugated anti-CRT monoclonal antibody ab196159 and FITC-conjugated anti-CD4 monoclonal antibody were added and incubated at 4 °C for 1 h. After washing with PBS (0.3% FBS), the cells were suspended in the same buffer with propidium iodide (1 µg/mL). Expression of CRT and CD4 was measured on the FACSAria II (BD Bioscience) and analyzed by Flowjo software (TreeStar Ashland, OR).
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).
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.
This product has been referenced in:
- Jacobson ME et al. Delivery of 5'-triphosphate RNA with endosomolytic nanoparticles potently activates RIG-I to improve cancer immunotherapy. Biomater Sci 7:547-559 (2019). Flow Cyt . Read more (PubMed: 30379158) »
- Ohkuro M et al. Calreticulin and integrin alpha dissociation induces anti-inflammatory programming in animal models of inflammatory bowel disease. Nat Commun 9:1982 (2018). Flow Cyt ; Human . Read more (PubMed: 29773794) »