Anti-Calcium Homeostasis Modul

Anti-Calcium Homeostasis Modulator 1-ATTO-594

CalHM1, FAM26C
Cat #: ACC-0101-AR
Sizes: 50 µl
Source: Rabbit
Type: Polyclonal
Applications: IC, IH
Reactivity: H, M, R

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, IC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IH- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Anti-Calcium Homeostasis Modulator 1 (#ACC-101) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize calcium homeostasis modulator 1 from rat, mouse, and human samples.

Alomone Labs is pleased to offer a new version of this antibody that is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-Calcium Homeostasis Modulator 1–ATTO-594 antibody (#ACC-101-AR) has been tested in immunohistochemistry applications and is specially suited for experiments requiring simultaneous labeling of different markers. The conjugated antibody is ideal for immuno-colocalization studies. For example we demonstrate that Calcium Homeostasis Modulator 1 co-localizes with GluN1 in mouse hippocampus using labeled CALHM1 antibody and Anti-NMDA Receptor (GluN1) (extracellular) antibody (#AGC-001).

• Applications
• Specifications
• Scientific Background
• Related Products

Immunohistochemistry

Expression of Calcium Homeostasis Modulator 1 in rat hippocampus
Immunohistochemical staining of rat hippocampus using Anti-Calcium Homeostasis Modulator 1-ATTO-594 antibody (#ACC-101-AR).  A. Staining of Calcium Homeostasis Modulator 1 appears only in the hippocampal CA1 pyramidal layer.  B. Nuclear staining using DAPI as the counterstain.  C. Merge of A and B.

Immuno-colocalization of GluN1 and CALHM1 in mouse hippocampal CA1 region
Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDA Receptor 1 (GluN1) (extracellular) antibody (#AGC-001), (1:200) and Anti-Calcium Homeostasis Modulator 1-ATTO-594 antibody (#ACC-101-AR), (1:60). A. Sections were stained with Anti-NMDA Receptor 1 (GluN1) (extracellular) antibody, followed by goat-anti-rabbit-Cy2 (green). Staining reveals expression in neurons of the pyramidal layer (an arrow points at the layer). B. The same section was incubated with Anti-Calcium Homeostasis Modulator 1-ATTO-594 antibody, (red). C. Merge of the two images demonstrates colocalization of GluN1 and CALHM1 in pyramidal neurons.
Immunogen
Peptide (C)EAMNHDLELGHTHG, corresponding to amino acid residues 252-265 of human calcium homeostasis modulator protein 1  (Accession Q8IU99).

HomologyRat, mouse - identical.
PurityAffinity purified on immobilized antigen.
FormulationLyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.
LabelATTO-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.
Standard quality control of each lotWestern blot analysis (unlabeled antibody, #ACC-101), and immunohistochemistry (labeled antibody).
Peptide confirmationConfirmed by amino acid analysis and mass spectrometry.
Storage before reconstitutionThe antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution50 µl double distilled water (DDW).
Antibody concentration after reconstitution0.8 mg/ml.
Storage after reconstitutionThe 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).
Control antigen storage before reconstitutionLyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
Control antigen reconstitution100 µl double distilled water (DDW).
Control antigen storage after reconstitution-20ºC.
Preadsorption Control1 µg peptide per 1 µg antibody.
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 metabolism^2,3. 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.

References

1. Green, K.N. and LaFerla, F.M. (2008) Neuron 59, 190.
2. Koppel, J. et al. (2011) Mol. Med. 17, 974.
3. Dreses-Werringloer, U. et al. (2008) Cell 133, 1149.