Immunofluorescence analysis of Calmodulin was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Calmodulin (6D4) Mouse Monoclonal Antibody (MA3918) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bacteria, Bovine, Chicken, Human, Rat|
|Published species reactivity||Rat, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Calmodulin purified from Dictyostelium discoideum.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||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.
MA3-918 detects calmodulin from bovine, chicken, Chlamydomonas, Dictyostelium, and rat samples. This antibody does not detect parvalbumin, tropinin, S-100, or myosin light chain kinase (MLCK).
MA3-918 has been successfully used in Western blot, immunocytochemistry and ELISA procedures. By Western blot, this antibody detects a 17 kDa protein representing calmodulin from Dictyostelium cell lysate. Addition of EGTA to buffers completely inhibits antibody staining. Immunohistochemical staining of calmodulin in Dictyostelium cells with MA3-918 results in staining of the contractile vacuoles.
The MA3-918 antigen is calmodulin purified from Dictyostelium discoideum.
Calmodulin is a small, highly conserved calcium binding protein found in all eukaryotic cells. With the capacity to bind up to four calcium ions, this 17 kDa protein acts as an important intracellular receptor for regulatory calcium signals. As it binds calcium, calmodulin undergoes conformational changes which can increase its affinity for target proteins. It acts both directly, through interaction with key target enzymes, and indirectly, via specific kinases. Studies have found that calmodulin participates in the regulation of several biological processes including energy and biosynthetic metabolism, cell motility, exocytosis, cytoskeletal assembly, and intracellular modulation of both cAMP and calcium concentrations.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Expression of calcium-buffering proteins in rat intrinsic laryngeal muscles.
MA3-918 was used in western blot to examine calcium-related proteins in the intrinsic laryngeal muscles, cricothyroid, and tibialis anterior muscles from male Sprague-Dawley rats
|Ferretti R,Marques MJ,Khurana TS,Santo Neto H||Physiological reports (3:null)||2015|
Regulation of sperm flagellar motility activation and chemotaxis caused by egg-derived substance(s) in sea cucumber.
MA3-918 was used in western blot to investigate the effect of egg-derived substances on sperm flagellar motility activation and chemotaxis
|Morita M,Kitamura M,Nakajima A,Sri Susilo E,Takemura A,Okuno M||Cell motility and the cytoskeleton (66:202)||2009|
Roles of calmodulin and calcium/calmodulin-dependent protein kinase in flagellar motility regulation in the coral Acropora digitifera.
MA3-918 was used in western blot to investigate the involvement of calmodulin and calcium/calmodulin-dependent protein kinase in the sperm flagellar motility regulation of Acropora Digitifera
|Morita M,Iguchi A,Takemura A||Marine biotechnology (New York, N.Y.) (11:118)||2009|
Increased sensitivity of the ryanodine receptor to halothane-induced oligomerization in malignant hyperthermia-susceptible human skeletal muscle.
MA3-918 was used in western blot to investigate the importance of RyR2 complex formation in the development of a metabolic crisis in malignant hyperthermia
|Glover L,Heffron JJ,Ohlendieck K||Journal of applied physiology (Bethesda, Md. : 1985) (96:11)||2004|
Characterization of the calcium-binding contractile protein centrin from Tetraselmis striata (Pleurastrophyceae).
MA3-918 was used in western blot to investigate the biochemical properties of centrin purified from Tetraselmis striata
|Coling DE,Salisbury JL||The Journal of protozoology (39:385)||1992|
Calcium-signaling components in rat insulinoma ß-cells (INS-1) and pancreatic islets are differentially influenced by melatonin.
MA3-918 was used in immunocytochemistry to study the differential effects of melatonin on the calcium signaling pathways of INS1 insulinoma cells and pancreatic islets
|Bazwinsky-Wutschke I,Mühlbauer E,Albrecht E,Peschke E||Journal of pineal research (56:439)||2014|