Overview

  • Product name
    Anti-Calreticulin antibody
    See all Calreticulin primary antibodies
  • Description
    Rabbit polyclonal to Calreticulin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC, ICC/IF, ELISA, IP, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Dog, Human, Drosophila melanogaster, Non human primates
  • Immunogen

    Other Immunogen Type corresponding to Human Calreticulin. Recombinant human calreticulin protein produced in the Baculovirus insect cell system.

  • Positive control
    • IHC: Rat brain cortex tissue: ICC: COS7 cells transfected with GFP-MAR2 or murine bone marrow macrophages infected with bacteria and fixed with 3% paraformaldehyde

Properties

Applications

Our Abpromise guarantee covers the use of ab2907 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 1/50 - 1/200.
ICC/IF Use at an assay dependent concentration. PubMed: 16943324
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/1000.
IHC-P Use at an assay dependent concentration. PubMed: 20731657
Flow Cyt Use at an assay dependent concentration. PubMed: 20388795

 ab171870, Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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
  • Alternative names
    • Autoantigen RO antibody
    • CALR antibody
    • CALR protein antibody
    • CALR_HUMAN antibody
    • Calregulin antibody
    • Calreticulin antibody
    • cC1qR antibody
    • CRP55 antibody
    • CRT antibody
    • CRTC antibody
    • Endoplasmic reticulum resident protein 60 antibody
    • Epididymis secretory sperm binding protein Li 99n antibody
    • ERp60 antibody
    • FLJ26680 antibody
    • grp60 antibody
    • HACBP antibody
    • HEL S 99n antibody
    • RO antibody
    • Sicca syndrome antigen A (autoantigen Ro; calreticulin) antibody
    • Sicca syndrome antigen A antibody
    • SSA antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of Calreticulin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2907 (1:75) for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 650 Phalloidin (1:120) and nuclei (blue) were stained with Hoechst (1ug/ml) for 30 minutes. Images were taken at 20X magnification.

  • All lanes : Anti-Calreticulin antibody (ab2907) at 1/1000 dilution

    All lanes : Whole cell lysate prepared from mouse skeletal muscle

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : HRP-conjugated mouse polyclonal to rabbit Ig at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 3 seconds

    See Abreview

  • ab2907 staining calreticulin in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 15% serum for 60 minutes at 20°C; antigen retrieval was by heat mediation in Tris/EDTA pH 9. Samples were incubated with primary antibody (1/500 in TBS) for 18 hours at 20°C. An Alexa Fluor® 647-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • ab2907 used at a 1/1000 dilution staining Calreticulin in HeLa cells by Immunocytochemistry/ Immunofluorescence.
    HeLa cells were transfected overnight with empty vector or plasmids encoding the indicated IFITM3 constructs. Immunofluorescence with a-HA antibodies allowed IFITM3 visualization, and a-calreticulin staining allowed visualization of the ER. TOPRO-3 was used to visualize nuclei. Scale bars indicate 10 µm. Ub? indicates mutation of Lys-24, Lys-83, Lys-88, and Lys-104 to alanine.
  • Immunocytochemistry/Immunofluorescence analysis of Calreticulin (red) in U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2907 (1:75) for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 633 goat anti-rabbit IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 488 Phalloidin (1:300) and nuclei (blue) were stained with Hoechst (1ug/ml) for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemsitry/Immunofluorescence analysis of Calreticulin (green) U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS 0.1% triton-X for 30 minutes at room temperature. Cells were incubated with ab2907 (1:50) for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin filaments (red) were stained with DyLight 554-Phalloidin (1:300) in PBS and incubated for 30 minutes. Nuclei (blue) were stained with Hoechst 33342 dye (1µg/mL). Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Calreticulin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2907 (1:75) for at least 1 hour at room temperature, washed with PBS, and incubated with Dylight 488 goat anti-rabbit IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with Dylight 350 Phalloidin (1:120) and nuclei (red) were stained with DRAQ5 (1ug/ml) for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Calreticulin (green) in A431 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2907 (1:75) for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin (1:120) and nuclei (blue) were stained with Hoechst (1ug/ml) for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of HMVEC cells labelling Calreticulin using ab2907.

  • Immunofluorescence analysis of HepG2 cells, staining Calreticulin with ab2907.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Saponin and blocked with 10% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/200 in PBS + 0.1% saponin) for 1 hour at 20°C. An AlexaFluor®647-conjugated donkey anti-rabbit polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Hossain DMS  et al. Dinaciclib induces immunogenic cell death and enhances anti-PD1-mediated tumor suppression. J Clin Invest 128:644-654 (2018). Read more (PubMed: 29337311) »
  • Anker JF  et al. Multi-faceted immunomodulatory and tissue-tropic clinical bacterial isolate potentiates prostate cancer immunotherapy. Nat Commun 9:1591 (2018). Read more (PubMed: 29686284) »
See all 99 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 25 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 24 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 2% · Temperature: 20°C
Sample
Fruit fly (Drosophila melanogaster) Cell (hemocyte (S2R+ cell))
Specification
hemocyte (S2R+ cell)
Permeabilization
Yes - 0.1% TX-100
Fixative
Paraformaldehyde

Miguel Sanchez

Verified customer

Submitted Jul 18 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HepG2)
Specification
HepG2
Fixative
Paraformaldehyde
Permeabilization
Yes - Saponin 0.1%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 30 2012

Question
Answer

Thank you for contacting Abcam yesterday.
The formulation for ab2907 should be 100 µl of antiserum containing 0.05% sodium azide. There is no PBS present.
I will have our datasheets team update the webpage as soon as possible.
I do apologize if this has created any issue.
Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you for contacting us.

The antibody ab2907 was tried in Arabidopsis by a customer, and the results were submitted as an Abreview (2 out of 5 stars): https://www.abcam.com/index.html?datasheet=2907&tab=abreviews&intabreviewid=24966

Thus, the Arabidopsis data is on the datasheet, but due to the rating of the results we do not list Arabidopsis as tested species or guarantee the antibody for it.

I have checked the homology of Arabidopsis with the human sequence and the homology is only 54% (isoform 1), 55% (isoform 2), and 49% (isoform 3) - thus very low. It is similar for the rabbit sequence: the homology between rabbit and Arabidopsis is also only 54%.
We usually suggest a sequence homology of 85% or higher forpotentialcross-reactivity. Thus, in generell it would be difficult to find an antibody raised against human or rabbit to be able to crossreact with Arabidopsis.

I might suggest to check http://www.biocompare.com for an antibody that was raised against an Arabidopsis sequence.

I hope this information is of some help to you. Please do not hesitate to contact us if you need any more advice or information.

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Arabidopsis thaliana Cell lysate - whole cell (cell culture)
Loading amount
20 µg
Specification
cell culture
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Joshua Heazlewood

Verified customer

Submitted Jul 14 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat1)
Specification
Rat1
Fixative
Paraformaldehyde
Permeabilization
Yes - NP40
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Timothy Angelotti

Verified customer

Submitted Mar 26 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (keratinocyte)
Specification
keratinocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton in PBS
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 16 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (skeletal muscle)
Loading amount
30 µg
Specification
skeletal muscle
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Oct 17 2008

1-10 of 16 Abreviews or Q&A

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