Name: Anti-CALHM2 (extracellular) Antibody
Brand: Alomone Labs
SKU: ACC-142

Overview

Cat #: ACC-142

Alternative Name Calcium homeostasis modulator protein 2, Protein FAM26B, FAM26B

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, m, r

Immunogen

Peptide (C)SEFVDPSSLTAGDK, corresponding to amino acid residues 133 - 146 of mouse CALHM2 (Accession Q8VEC4). Extracellular, 2^nd loop.

Accession (Uniprot) Number Q8VEC4

Gene ID 72691

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Rat – 13 out of 14 amino acid residues identical. Human - 11 out of 14 amino acid residues identical.

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4.

Isotype Rabbit IgG

Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.

Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.

Reconstitution 0.2 ml double distilled water (DDW).

Antibody concentration after reconstitution 0.8 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C 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 x g 5 min).

Preadsorption Control 1 µg peptide per 1 µg antibody
CALHM2 (extracellular) Blocking Peptide (#BLP-CC142)

Standard quality control of each lot Western blot analysis.

Applications: ifc, ih, lci, wb

May also work in: ic*, ip*

Western blot

Western blot analysis of mouse brain lysate (lanes 1 and 3) and rat brain lysate (lanes 2 and 4):
1-2. Anti-CALHM2 (extracellular) Antibody (#ACC-142), (1:200).
3-4. Anti-CALHM2 (extracellular) Antibody, preincubated with CALHM2 (extracellular) Blocking Peptide (BLP-CC142).
Western blot analysis of human HMC3 microglia cell line lysate (lanes 1 and 3) and human U-87 MG glioma cell line lysate (lanes 2 and 4):
1-2. Anti-CALHM2 (extracellular) Antibody (#ACC-142), (1:200).
3-4. Anti-CALHM2 (extracellular) Antibody, preincubated with CALHM2 (extracellular) Blocking Peptide (BLP-CC142).

Immunohistochemistry

Expression of CALHM2 in rat spinal cord.
Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections with Anti-CALHM2 (extracellular) Antibody (#ACC-142), (1:300), followed by goat anti-rabbit-AlexaFluor-488. A. CALHM2 immunoreactivity (green) appears in layers 1-2 of the dorsal horn (vertical arrows) and in some nerve cells (horizontal arrows). B. Pre-incubation of the antibody with CALHM2 (extracellular) Blocking Peptide (BLP-CC142), suppressed staining. Cell nuclei are stained with DAPI (blue).

Indirect flow cytometry

Cell surface detection of CALHM2 by indirect flow cytometry in live intact mouse BV-2 microglia cell line:
^___ Cells.
^___ Cells + goat-anti-rabbit-FITC.
^___ Cells + Anti-CALHM2 (extracellular) Antibody (#ACC-142), (2.5μg) + goat-anti-rabbit-FITC.

References

Ma, Z. et al. (2016) Pflugers. Archiv. Eur. J. Physiol. 468,395.
Syrjanen, J.L. et al. (2020) Nat. Struct. Mol. Biol. 27,2.
Choi, W. et al. (2019) Nature. 576,7785.
Cheng, J. et al. (2021) Sci. Adv. 7, eabe3600.

Scientific background

Calcium homeostasis modulator protein 2 (CALHM2) is a member of the CALHM channel family, a group of transmembrane proteins that play essential roles in calcium ion transport and homeostasis. The CALHM family consist in humans of six members: CALHM1 to CALHM6.¹

CALHM2 functions as a calcium-permeable ion channel. It allows calcium ions to pass through the cell membrane in response to specific stimuli, such as changes in membrane potential or extracellular calcium concentration.^2,3

CALHM2 is composed of 323 amino acids and assembles as an undecamer with each protomer containing a large N-terminal transmembrane (TM) domain with 4 TM helices, an intracellular C-terminal domain, and a small extracellular linker region. ^2,3

CALHMs permeate ions and ATP in a voltage-dependent manner to modulate neuronal excitability. CALHM2 has been shown to accommodate lipids in its hydrophobic channel in order to regulate channel function.^2,3

Ion and ATP permeation by CALHM proteins play a role in the physiology of cognition, depression, gustatory signaling, and the pathology of Alzheimer’s disease.

Specifically, CALHM2 expression was increased in Alzheimer's disease (AD) patients and in an AD model in mice. Knockout of CALHM2 improved Aβ plaque deposition, reduced neuroinflammation, and improved cognitive impairment in an AD mice model.⁴

Moreover, microglial CALHM2 may have a central role in both chronic inflammatory diseases (like AD) and in acute inflammatory reactions and neuroinflammation.⁴

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

Species reactivity key:

H- Human, M- Mouse, R- Rat

Lyophilized Powder

Anti-CALHM2 (extracellular) Antibody (#ACC-142) is a highly specific antibody directed against an extracellular epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry and flow cytometry applications. It has been designed to recognize CALHM2 from mouse, rat and human samples.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 29/10/2024

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Anti-CALHM2 (extracellular) Antibody

Overview

Application key:

Species reactivity key:

Applications

Citations

Specifications

Scientific Background

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