Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #ACC-101), and immunohistochemistry (labeled antibody).
Applications: ic, if, ih
Immunohistochemistry
Rat and mouse hippocampus frozen sections (1:100).
Expression of CALHM1 in rat hippocampus
Immunohistochemical staining of rat hippocampus using Anti-CALHM1-ATTO Fluor-594 Antibody (#ACC-101-AR). A. Staining of CALHM1 appears only in the hippocampal CA1 pyramidal layer. B. Nuclear staining using DAPI as the counterstain. C. Merge of A and B.
Scientific background
A misbalance in Ca2+ homeostasis seems to be related to the development of Alzheimer’s disease (AD). Indeed Neuronal Ca2+ balance may affect the levels of proteins associated with AD, such as amyloid-beta (Ab) and tau1.
Calcium homeostasis modulator 1 (CALHM1) is a cerebral Ca2+ channel component responsible for controlling intracellular Ca2+ levels and Ab metabolism2,3. CALHM1 is a three transmembrane glycoprotein, mostly localized to the endoplasmic reticulum (ER) although it can be detected at the plasma membrane. A functional channel is formed by multiple subunits which has some structural similarities to the NMDA receptor. It is expressed in all brain regions1.
In cultured cells, overexpression of CALHM1 increases intracellular Ca2+ levels and reduces Ab accumulation, thereby reinforcing its implication in the development of AD. Indeed, a polymorphism in the gene was found to affect the onset of AD2.
Application key:
CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot
Expression of CALHM1 in rat hippocampusImmunohistochemical staining of rat hippocampus using Anti-CALHM1-ATTO Fluor-594 Antibody (#ACC-101-AR). A. Staining of CALHM1 appears only in the hippocampal CA1 pyramidal layer. B. Nuclear staining using DAPI as the counterstain. C. Merge of A and B.