Recombinant Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)
![Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)](https://www.abcam.com/ps/products/196/ab196158/Images/ab196158-286075-anti-calreticulin-antibody-epr3924-er-marker-alexa-fluor-488-immunofluorescence.jpg "Immunocytochemistry/ Immunofluorescence - 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: ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
- Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm
Related conjugates and formulations
Overview
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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: ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Monkey
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab180826) -
Positive control
- ICC/IF: HeLa cells, HAP1 cells (HAP1-CALR knockout cells used as negative cell line) Flow Cyt (intra): HeLa cells, 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... -
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® 647 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196159)
- 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 ab196158 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 |
1/1000.
This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min) |
|
| Flow Cyt (Intra) |
1/50.
ab199091 - Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
ICC/IF
1/1000. This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min) |
|
Flow Cyt (Intra)
1/50. ab199091 - Rabbit monoclonal IgG (Alexa Fluor® 488), is suitable for use as an isotype control with this antibody. |
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® 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).
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![Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)](https://www.abcam.com/ps/products/196/ab196158/Images/ab196158-280188-anti-calreticulin-antibody-epr3924-er-marker-alexa-fluor-488-immunofluorescence.jpg)
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).
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![Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)](https://www.abcam.com/ps/products/196/ab196158/Images/ab196158-296962-anti-calreticulin-antibody-epr3924-er-marker-alexa-fluor-488-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - 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. -
![Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)](https://www.abcam.com/ps/products/196/ab196158/Images/ab196158-239566-anti-calreticulin-antibody-epr3924-er-marker-alexa-fluor-488-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - 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.
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![Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)](https://www.abcam.com/ps/products/196/ab196158/Images/ab196158-7-benefits-of-recombinant-antibodies.png)
Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (16)
ab196158 has been referenced in 16 publications.
- Li Y et al. Targeted immunotherapy for HER2-low breast cancer with 17p loss. Sci Transl Med 13:N/A (2021). PubMed: 33568521
- Huang KC et al. ATAD3A stabilizes GRP78 to suppress ER stress for acquired chemoresistance in colorectal cancer. J Cell Physiol N/A:N/A (2021). PubMed: 33580514
- Arioka Y et al. Chromosome 22q11.2 deletion causes PERK-dependent vulnerability in dopaminergic neurons. EBioMedicine 63:103138 (2021). PubMed: 33341442
- Huang Z et al. Nanoscale coordination polymers induce immunogenic cell death by amplifying radiation therapy mediated oxidative stress. Nat Commun 12:145 (2021). PubMed: 33420008
- Flieswasser T et al. Clinically Relevant Chemotherapeutics Have the Ability to Induce Immunogenic Cell Death in Non-Small Cell Lung Cancer. Cells 9:N/A (2020). PubMed: 32560232