Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker (Alexa Fluor® 488)
- Suitable for: Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
- Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm
Product nameAnti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 488)
See all Calreticulin primary antibodies
DescriptionRabbit monoclonal [EPR3924] to Calreticulin - ER Marker (Alexa Fluor® 488)
ConjugationAlexa Fluor® 488. Ex: 495nm, Em: 519nm
Tested applicationsSuitable for: Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Monkey
Synthetic peptide within Human Calreticulin aa 50-150. The exact sequence is proprietary.
(Peptide available as
- ICC/IF: HeLa cells, HAP1 cells (HAP1-CALR knockout cells used as negative cell line) Flow Cyt: HeLa cells, 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 instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
PurityProtein A purified
- Anti-Calreticulin antibody [EPR3924] - ER Marker (HRP) (ab195511)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (Alexa Fluor® 647) (ab196159)
- Anti-Calreticulin antibody [EPR3924] - ER Marker (PE) (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)
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab196158 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab199091 - Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control with this antibody.
This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min)
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
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).
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).
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.
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).
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.
ab196158 has been referenced in 10 publications.
- Lin A et al. Non-Thermal Plasma as a Unique Delivery System of Short-Lived Reactive Oxygen and Nitrogen Species for Immunogenic Cell Death in Melanoma Cells. Adv Sci (Weinh) 6:1802062 (2019). PubMed: 30937272
- Chaurasiya S et al. A chimeric poxvirus with J2R (thymidine kinase) deletion shows safety and anti-tumor activity in lung cancer models. Cancer Gene Ther N/A:N/A (2019). Flow Cyt ; Mouse, Human, African green monkey . PubMed: 31209267
- Van Loenhout J et al. Cold Atmospheric Plasma-Treated PBS Eliminates Immunosuppressive Pancreatic Stellate Cells and Induces Immunogenic Cell Death of Pancreatic Cancer Cells. Cancers (Basel) 11:N/A (2019). PubMed: 31635070
- Guo S et al. Nano-pulse stimulation induces potent immune responses, eradicating local breast cancer while reducing distant metastases. Int J Cancer 142:629-640 (2018). PubMed: 28944452
- Lin SY et al. Necroptosis promotes autophagy-dependent upregulation of DAMP and results in immunosurveillance. Autophagy 14:778-795 (2018). PubMed: 29171784
- O'Leary MP et al. Novel oncolytic chimeric orthopoxvirus causes regression of pancreatic cancer xenografts and exhibits abscopal effect at a single low dose. J Transl Med 16:110 (2018). PubMed: 29699566
- Pan W et al. A novel SMAC mimetic APG-1387 exhibits dual antitumor effect on HBV-positive hepatocellular carcinoma with high expression of cIAP2 by inducing apoptosis and enhancing innate anti-tumor immunity. Biochem Pharmacol 154:127-135 (2018). Human . PubMed: 29679556
- Zhang Q et al. Sonodynamic therapy-assisted immunotherapy: A novel modality for cancer treatment. Cancer Sci 109:1330-1345 (2018). PubMed: 29575297
- Zhang Y et al. Enhancing CD8+ T Cell Fatty Acid Catabolism within a Metabolically Challenging Tumor Microenvironment Increases the Efficacy of Melanoma Immunotherapy. Cancer Cell 32:377-391.e9 (2017). PubMed: 28898698
- Ng HK et al. Signaling Modification by GPCR Heteromer and Its Implication on X-Linked Nephrogenic Diabetes Insipidus. PLoS One 11:e0163086 (2016). PubMed: 27649563