Recombinant Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
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
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
-
Product name
Anti-Calreticulin antibody [EPR3924] - ER Marker
See all Calreticulin primary antibodies -
Description
Rabbit monoclonal [EPR3924] to Calreticulin - ER Marker -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Monkey -
Immunogen
-
Positive control
- WB: SH-SY5Y, HL-60, HepG2, HeLa, Fetal kidney and Fetal brain lysates; Human kidney tissue; Mouse and Rat brain lysates. ICC/IF: HAP1 cells (HAP1-CALR as negative cell line) IHC-P: Human colon, kidney, liver, placenta, stomach, breast carcinoma and Papillary carcinoma of thyroid gland tissues; Mouse liver and Rat lung tissues.
-
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3924 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
- HRP Anti-Calreticulin antibody [EPR3924] - ER Marker (ab195511)
- Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab196158)
- 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)
- Anti-Calreticulin antibody [EPR3924] - BSA and Azide free (ab271865)
- Alexa Fluor® 555 Anti-Calreticulin antibody [EPR3924] (ab275117)
- Alexa Fluor® 594 Anti-Calreticulin antibody [EPR3924] - ER Marker (ab275343)
-
Compatible Secondaries
-
Immunizing Peptide (Blocking)
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
Applications
Our Abpromise guarantee covers the use of ab92516 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 |
---|---|---|
WB | 1/1000 - 1/10000. Predicted molecular weight: 48 kDa.Can be blocked with Calreticulin peptide (ab180826). | |
IHC-P | 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The use of a HRP/AP polymerized secondary antibody is recommended for enhanced staining. |
|
Flow Cyt | 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | 1/500. |
Target
-
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
-
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
-
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Calreticulin knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: NIH3T3 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab92516 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab92516 was shown to specifically react with Calreticulin when Calreticulin knockout samples were used. Wild-type and Calreticulin knockout samples were subjected to SDS-PAGE. ab92516 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
-
Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
ab92516 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 ab92516 at 1/500 and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 488 (ab196158) and Alexa Fluor® 647 (ab196159) conjugated versions are available for this clone.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat lung.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
-
All lanes : Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516) at 1/10000 dilution
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) Whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : Mouse brain lysates
Lane 4 : Rat brain lysates
Lane 5 : COS-1 (African green monkey kidney fibroblast-like) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane 1 to 3: 10 seconds; Lane 4 and 5: 130 seconds
-
Overlay histogram showing HAP1 wildtype (green line) and HAP1-CALR knockout cells (red line) stained with ab92516. 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 (ab92516, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.
A rabbit IgG isotype control antibody (ab172730) 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 50 mW Blue laser (488nm) and 530/30 bandpass filter.Alexa Fluor® 488 (ab196158) and Alexa Fluor® 647 (ab196159) conjugated versions are available for this clone.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on mouse liver.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Calreticulin with ab92516, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining human breast carcinoma.The section was incubated with ab229902 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Formalin-fixed, paraffin-embedded normal human colon tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunocytochemistry/ Immunofluorescence - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) labelling Calreticulin with purified ab92516 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
-
Overlay histogram showing HeLa cells stained with ab92516 (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 (ab92516, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
-
All lanes : Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516) at 1/1000 dilution
Lane 1 : SH-SY5Y cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : Human fetal kidney lysate
Lane 6 : Human fetal brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 48 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
ab92516, at 1/250 dilution, staining Calreticulin in paraffin embedded Human kidney tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
ab92516 showing negative staining in Normal human heart tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Formalin-fixed, paraffin-embedded normal human liver tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Formalin-fixed, paraffin-embedded normal human placenta tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Formalin-fixed, paraffin-embedded normal human stomach tissue stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calreticulin antibody [EPR3924] - ER Marker (ab92516)
Formalin-fixed, paraffin-embedded Papillary carcinoma of human thyroid gland tissue.stained for Calreticulin using ab92516 at 1/250 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
References (36)
ab92516 has been referenced in 36 publications.
- Eckhardt J et al. Molecular basis of impaired extraocular muscle function in a mouse model of congenital myopathy due to compound heterozygous Ryr1 mutations. Hum Mol Genet 29:1330-1339 (2020). PubMed: 32242214
- Wolf-Dennen K et al. Exosomal communication by metastatic osteosarcoma cells modulates alveolar macrophages to an M2 tumor-promoting phenotype and inhibits tumoricidal functions. Oncoimmunology 9:1747677 (2020). PubMed: 32313728
- Wan H et al. WDR45 contributes to neurodegeneration through regulation of ER homeostasis and neuronal death. Autophagy 16:531-547 (2020). PubMed: 31204559
- Singhal A et al. 2-Hydroxypropyl-gamma-cyclodextrin overcomes NPC1 deficiency by enhancing lysosome-ER association and autophagy. Sci Rep 10:8663 (2020). PubMed: 32457374
- Yang C et al. Nano-encapsulated tryptanthrin derivative for combined anticancer therapy via inhibiting indoleamine 2,3-dioxygenase and inducing immunogenic cell death. Nanomedicine (Lond) 14:2423-2440 (2019). PubMed: 31549585