Immunofluorescent analysis of Calreticulin (green) in SK-BR-3 cells. The cells were fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a Calreticulin mouse monoclonal antibody (Product # MA5-15382) at a concentration of 10ug/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). F-actin (red) was stained by Dylight 554 Phalloidin (Product #21834) and nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Synthetic peptide corresponding to aa (EEEDVPGQAKDELC) of human Calreticulin, conjugated to KLH.|
|Contains||0.05% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1-3 µg/ml|
|Immunocytochemistry (ICC)||5-10 µg/ml|
|Immunofluorescence (IF)||5-10 µg/ml|
|Immunohistochemistry (IHC)||1:200 - 1:1000|
|Western Blot (WB)||2 µg/ml|
* 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.
MA5-15382 targets Calreticulin in IF, IHC, and WB applications and shows reactivity with Human and mouse samples.
The MA5-15382 immunogen is synthetic peptide corresponding to aa (EEEDVPGQAKDELC) of human Calreticulin, conjugated to KLH.
MA5-15382 detects Calreticulin which has a predicted molecular weight of approximately 48kDa.
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.
Serum calreticulin is a negative biomarker in patients with Alzheimer's disease.
MA5-15382 was used in western blot to measure calreticulin in patients with Alzheimer's disease and assess it's use as a biomarker for disease
|Lin Q,Cao Y,Gao J||International journal of molecular sciences (15:21740)||2014|
Platelet protein disulfide isomerase is localized in the dense tubular system and does not become surface expressed after activation.
MA5-15382 was used in immunohistochemistry to study the subcellular localization of platelet protein disulfide isomerase
|van Nispen Tot Pannerden HE,van Dijk SM,Du V,Heijnen HF||Blood (114:4738)||2009|