Anti-Calreticulin / CALR antibody, rabbit serum
Product code73-018

Size

100 µlStorage

Store 4℃ for short term For long term storage store at -20℃.
Aliquot to avoid repeated freezing and thawing.

Concentration

N/A

Buffer

0.09% sodium azide

PurityRabbit antiserum

Immunogen

C-EEDEKEEDEEESPGQAKDEL (C-terminal of mouse CALR protein)

IsotypeRabbit IgG

Reactivity

Mouse and human.
Not tested with other species.

Special notes

Validation: Knock-out mouse

Application

1. Western blotting (1/500~1/1,000 dilution))
2. Immunoprecipitation (1/100)
3. Immunoflourescence staining (1/300~1/1,000 dilution)
4. Immunohistochemistry (1/1,000 dilution)
Other applications have not been tested.

Background

Calreticulin (CALR) is calcium-binding chaperone that promotes folding,oligomeric assembly and quality control in the endoplasmic reticulum (ER) via
the calreticulin/calnexin cycle. This lectin interacts transiently with almost allof the monoglucosylated glycoproteins that are synthesized in the ER. Interacts
with the DNA-binding domain of NR3C1 and mediates its nuclear export.
Involved in maternal gene expression regulation. May participate in oocyte
maturation via the regulation of calcium homeostasis

Data Link

uniprot/P14211 Mouse Calrecticulin, uniprot/Q96L12 Human Calreticulin
Gene I D 12317 Mouse Calr,

Gene ID 811 Human Calr

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 73-018 Anti-Calreticulin / CALR antibody, rabbit serum

Fig.1 Identification of CALR protein by western blotting with anti-CALR antibody.
Embryonic fibroblast cells prepared from Calr -/- mouse were transfected with a plasmid expressing

Calr. The cell lysate was analyzed by western blotting with anti-CALR antibody at 1/500 dilution.
1. Mock-infected cell lysate as a negative control.
2. Cell lysate transfected with a plasmid expressing Calr.
The molecular mass is 48 kDa

Fig.2 Western blot analysis of CALR protein in crude extracts of mouse testis and sperm with antiCALR antibody.
Proteins in the extracts (10 μg protein) were separated on 10-20% gradient gel of SDS-PAGE andelectro-blotted to a PVDF membrane. The membrane was reacted with anti-CALR antibody at 1/1,000dilution. As the 2nd antibody, anti-rat IgG antibody conjugated with HRP (ab97051) was used at
1/10,000 dilution
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Fig.3 Western blot analysis of CALR protein in crude extracts of human cell lines with anti-CALRantibody.
Proteins in the extracts (10 μg protein) were separated on 12.5% gel of SDS-PAGE and electro-blotted
to a PVDF membrane (wet system). The membrane was reacted with anti-CALR antibody at 1/1,000dilution. As the 2nd antibody, anti-rat IgG antibody conjugated with HRP (ab97051) was used at
1/10,000 dilution.

Fig.4. Immunoprecipitation of CALR protein with anti-CALR antibody.
Lysates of wild-type mouse testis were immunoprecipitated with anti-CALR antibody and theprecipitates were analyzed by western blotting with the same antibody.
1. Input testis lysate
2. Precipitated with preimmune serum
3. Precipitated with anti-CALR antibody

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Fig.5 Immunofluorescence staining of CALR protein in NIH3T3 cells with anti-CALR antibody.
NIH3T3 cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% TritonX 100 andreacted with anti-CALR antibody at 1/300 dilution. As the 2nd antibody, goat anti-rabbit IgG antibody
conjugated with Alexa Fluor 488 (Molecular Probes) was used at 1/1,000 dilution.
DNA was stained with DAPI (1 ug/ml) and the merged image was shown on right.

Fig.6. Immunohistological staining of CALR protein in mouse testis using anti-CALR antibody.
A section of formalin fixed and paraffin embedded mouse testis was treated with the anti-CALR
antibody at 1/1,000 dilution after deparaffization and antigen retrieval. The 2 nd antibody, goat antirabbit IgG conjugated with Alexa Fluor 488 (Molecular Probes #1166843) was used at 1/1,000 dilution.
DNA was stained with DAPI (1.0μg/mL) and the merged image was shown (Merge). The bright-field
image of the same region was shown on the right.
Reference: This antibody was described and used in the following publication.
1. Ikawa M. et al (2011) Calsperin is a testis-specific chaperone required for sperm fertility. J Biol
Chem.18:5639-46. pubmed/21131354 Free article. WB, IP

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