Western blot analysis was performed on membrane enriched extracts (30 ug lysate) of MCF7 (Lane 1), MDA-MB-231 (Lane 2), HeLa (Lane 3), K-562 (Lane 4), Jurkat (Lane 5) and A549 (Lane 6). The blot was probed with Chicken Anti-Calreticulin Polyclonal Antibody (Product # PA1-903, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Chicken IgY (H+L) Secondary Antibody, HRP conjugate (Product # A16054, 0.4 µg/ml, 1:2500 dilution). A 55 kDa band corresponding to Calreticulin was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product# SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Dog, Hamster, Human, Mouse, Rabbit, Rat|
|Published species reactivity||Rabbit, Rat, Amphibian, Human, Mouse|
|Host / Isotype||Chicken / IgY|
|Immunogen||Synthetic Peptide: D(399) E K E E D E E E S P G Q A K D(414).|
|Purification||Antigen affinity chromatography|
|Storage buffer||1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay dependent|
|Immunofluorescence (IF)||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:2,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-903 detects calreticulin from human, rat, mouse, hamster, rabbit and canine tissues.
PA1-903 has been successfully used in Western blot, immunoprecipitation, immunocytochemistry and immunofluorescence procedures. By Western blot, this antibody detects an ~60 kDa protein representing calreticulin and an ~120 kDa unidentified protein from rat brain extract.
PA1-903 immunizing peptide corresponds to amino acid residues 399-414 from mouse calreticulin. PA1-903 immunizing peptide (Cat. # PEP-017) is available for use in neutralization and control experiments.
Calreticulin is the major calcium binding protein found in smooth muscle sarcoplasmic reticulum (SR) and non-muscle endoplasmic reticulum (ER) membranes. This protein was originally identified in SR membranes and plays a minor role in calcium storage in skeletal and cardiac muscle SR. Calreticulin is also known as calregulin, CRP55, CaBP3, calsequestrin-like protein and Ro/SS-A antigen. Calreticulin binds calcium with low affinity and high capacity, however it also exhibits a single high affinity binding site. The highly conserved sequence Lys-Asp-Glu-Leu (KDEL) is present at the C-terminus of calreticulin and other resident ER proteins including glucose regulated protein 78 (GRP78), GRP94 and protein disulfide isomerase (PDI). This sequence is responsible for the retention of newly synthesized proteins within the ER lumen. This retention is reported to be mediated by a KDEL receptor. Recent reports indicate that calreticulin can act as a modulator of the regulation of gene transcription by nuclear hormone receptors and may also act as a molecular chaperone.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Bilateral acute pyogenic conjunctivitis with iritis induced by unilateral topical application of bacterial peptidoglycan muramyl dipeptide in adult rabbits.
PA1-903 was used in immunohistochemistry to study the role of bacterial peptidoglycan muramyl dipeptide in the pathogenesis of acute pyogenic conjunctivitis and iritis
|Langford MP,Foreman BD,Srur L,Ganley JP,Redens TB||Experimental eye research (116:324)||2013|
mTOR-Controlled Autophagy Requires Intracellular Ca(2+) Signaling.
PA1-903 was used in western blot to study the role of intracellular Ca(2+) signaling in mTOR-modulated autophagy
|Decuypere JP,Kindt D,Luyten T,Welkenhuyzen K,Missiaen L,De Smedt H,Bultynck G,Parys JB||PloS one (8:null)||2013|
Gestational exposure to diethylstilbestrol alters cardiac structure/function, protein expression and DNA methylation in adult male mice progeny.
PA1-903 was used in western blot to study the effects of diethylstilbestrol exposure during gestation on epigenetic DNA modifications and cardiac structure and function in adult mice
|Haddad R,Kasneci A,Mepham K,Sebag IA,Chalifour LE||Toxicology and applied pharmacology (266:27)||2013|
Comparative proteomics profiling of a phospholamban mutant mouse model of dilated cardiomyopathy reveals progressive intracellular stress responses.
PA1-903 was used in western blot to perform the global proteomics surveys of cardiac ventricle of a phospholamban mutant mouse
|Gramolini AO,Kislinger T,Alikhani-Koopaei R,Fong V,Thompson NJ,Isserlin R,Sharma P,Oudit GY,Trivieri MG,Fagan A,Kannan A,Higgins DG,Huedig H,Hess G,Arab S,Seidman JG,Seidman CE,Frey B,Perry M,Backx PH,Liu PP,MacLennan DH,Emili A||Molecular and cellular proteomics : MCP (7:519)||2008|
Cardiac mitochondrial connexin 43 regulates apoptosis.
PA1-903 was used in western blot to investigate the role of cardiac mitochondrial connexin 43 in apoptosis
|Goubaeva F,Mikami M,Giardina S,Ding B,Abe J,Yang J||Biochemical and biophysical research communications (352:97)||2007|
Localization of the iron transport proteins Mobilferrin and DMT-1 in the duodenum: the surprising role of mucin.
PA1-903 was used in western blot to study the intestinal iron uptake and the involvement of mucin
|Simovich M,Hainsworth LN,Fields PA,Umbreit JN,Conrad ME||American journal of hematology (74:32)||2003|
Interactions of HLA-B27 with the peptide loading complex as revealed by heavy chain mutations.
PA1-903 was used in western blot to investigate how HLA-B27 interact with the peptide loading complex
|Harris MR,Lybarger L,Myers NB,Hilbert C,Solheim JC,Hansen TH,Yu YY||International immunology (13:1275)||2001|
Association of ERp57 with mouse MHC class I molecules is tapasin dependent and mimics that of calreticulin and not calnexin.
PA1-903 was used in western blot to study the interaction between ERp57 and mouse MHC class I molecules.
|Harris MR,Lybarger L,Yu YY,Myers NB,Hansen TH||Journal of immunology (Baltimore, Md. : 1950) (166:6686)||2001|
Chaperone function of calreticulin when expressed in the endoplasmic reticulum as the membrane-anchored and soluble forms.
PA1-903 was used in western blot to demonstrate that calreticulin in ER is another novel CAS-sensitive chaperone
|Wada I,Imai S,Kai M,Sakane F,Kanoh H||The Journal of biological chemistry (270:20298)||1995|
C1q and mannose binding lectin engagement of cell surface calreticulin and CD91 initiates macropinocytosis and uptake of apoptotic cells.
PA1-903 was used in immunocytochemistry to study the mechanisms of macropinocytosis and apoptotic cell removal
|Ogden CA,deCathelineau A,Hoffmann PR,Bratton D,Ghebrehiwet B,Fadok VA,Henson PM||The Journal of experimental medicine (194:781)||2001|
Calreticulin inhibits repetitive intracellular Ca2+ waves.
PA1-903 was used in immunocytochemistry to investigate the role of CRT in IP3-mediated caclium signaling
|Camacho P,Lechleiter JD||Cell (82:765)||1995|