Product nameAnti-Calreticulin antibody [FMC 75]
See all Calreticulin primary antibodies
DescriptionMouse monoclonal [FMC 75] to Calreticulin
Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human, Monkey
Predicted to work with: Chinese hamster
Fusion protein corresponding to Calreticulin. Calreticulin-maltose binding fusion protein.
- WB: HeLa whole cell lysate. IHC-P: Human placenta tissue. ICC/IF: HeLa cell lysate. Flow: HeLa cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: PBS, pH 7.2
Concentration information loading...
Purification notesPurified from ascites.
Clone numberFMC 75
Light chain typekappa
Our Abpromise guarantee covers the use of ab22683 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Predicted molecular weight: 63 kDa.|
|IP||Use a concentration of 12.5 µg/ml.|
|Flow Cyt||Use 1-2µg for 106 cells.
(Additional resource: PMID - 18689689)
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Fix and permeabilize cells prior to staining.
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
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
Anti-Calreticulin antibody [FMC 75] (ab22683) at 1/2000 dilution + HEK293 whole cell lysate at 10 µg
HRP-conjugated goat anti-mouse IgG (H+L) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 42,65 kDa why is the actual band size different from the predicted?
Additional bands at: 30 kDa (possible non-specific binding)
Exposure time: 10 seconds
Blocked with 5% milk for 1 hour at 23°C.
Incubated with the primary antibody in PBS + 0.5% Tween 20 for 12 hours at 4°C.
ICC/IF image of ab22683 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22683, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Ab22683 staining human placenta. Staining is localised to the cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Overlay histogram showing HeLa cells stained with ab22683 (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 (ab22683, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This product has been referenced in:
- Zhao D et al. Cardiac-derived CTRP9 protects against myocardial ischemia/reperfusion injury via calreticulin-dependent inhibition of apoptosis. Cell Death Dis 9:723 (2018). Read more (PubMed: 29925877) »
- Feng M et al. Programmed cell removal by calreticulin in tissue homeostasis and cancer. Nat Commun 9:3194 (2018). Read more (PubMed: 30097573) »