asic1a (Alomone Labs)


Structured Review

Asic1a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/asic1a/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Demonstration of a Direct Interaction between σ-1 Receptors and Acid-Sensing Ion Channels"
Article Title: Demonstration of a Direct Interaction between σ-1 Receptors and Acid-Sensing Ion Channels
Journal: Biophysical Journal
doi: 10.1016/j.bpj.2009.12.4293

Figure Legend Snippet: Analysis of σ -1 receptor binding to ASIC1a. ( A ) Gallery of zoomed images of ASICs that are undecorated ( top ) or decorated by one ( middle ) or two ( bottom ) peripheral particles. Lines indicate the angles between pairs of bound peripheral particles. ( B–D ) Frequency distributions of ( B ) molecular volumes of decorated central particles, ( C ) molecular volumes of bound peripheral particles, and ( D ) angles between pairs of bound peripheral particles. The curves indicate the fitted Gaussian functions. The means of the distributions are indicated.
Techniques Used: Binding Assay

Figure Legend Snippet: AFM imaging of ASIC1a, anti-His 6 antibodies, and σ -1 receptors. ( A ) Low-magnification image of isolated ASIC1a. The arrow indicates a trimeric cluster of three equally sized particles, which is likely an ASIC trimer that has bound to the mica intact and then fallen apart during the drying process. A zoomed image of this cluster is shown at the bottom right of the panel. A shade-height scale is shown at the right. ( B ) Sections through the two particles indicated in A , taken at the positions indicated by the lines. The sections indicate that the shapes of the particles approximate that of a spherical cap. The calculated volumes of the particles are indicated. ( C ) Low-magnification image of anti-His 6 antibodies. A shade-height scale is shown at the right. ( D ) Frequency distribution of molecular volumes of anti-His 6 antibodies. The curve indicates the fitted Gaussian function. The mean of the distribution is indicated. ( E ) Low-magnification image of σ -1 receptors. A shade-height scale is shown at the right. ( F ) Frequency distribution of molecular volumes of σ -1 receptors. The curve indicates the fitted Gaussian function. The mean of the distribution is indicated.
Techniques Used: Imaging, Isolation

Figure Legend Snippet: Analysis of anti-His 6 antibody binding to His 8 -tagged ASIC1a. ( A ) Low-magnification AFM images of a sample of isolated ASIC1a that had been incubated with anti-His 6 antibody. Arrowheads indicate singly decorated ASICs; arrows indicate doubly decorated ASICs. A shade-height scale is shown at the right. ( B ) Gallery of zoomed images of ASICs that are undecorated ( top ) or decorated by one ( middle ) or two ( bottom ) anti-His 6 antibodies. Lines indicate the angles between pairs of bound antibodies. ( C–E ) Frequency distributions of ( C ) molecular volumes of decorated central particles, ( D ) molecular volumes of bound peripheral particles, and ( E ) angles between pairs of bound peripheral particles. The curves indicate the fitted Gaussian functions. The means of the distributions are indicated.
Techniques Used: Binding Assay, Isolation, Incubation

Figure Legend Snippet: Isolation of proteins from ASIC1a-expressing cells. ( A ) Immunofluorescence detection of ASIC1a in stably transfected HEK-293 cells. Cells were fixed, permeabilized, and incubated with either rabbit polyclonal anti-ASIC1a antibody or mouse monoclonal anti-His 6 antibody, followed by Cy3-conjugated goat anti-rabbit or anti-mouse secondary antibodies, as appropriate. Cells were imaged by confocal laser scanning microscopy. ( B ) Immunofluorescence detection of the σ -1 receptor in HEK-293 cells stably expressing ASIC1a and transiently transfected with σ -1 receptor cDNA. Cells were incubated with mouse monoclonal anti-FLAG antibody, followed by fluorescein isothiocynate-conjugated goat anti-mouse secondary antibodies. A representative immunofluorescence image is shown, along with a corresponding brightfield image; ∼20% of the cells are expressing the σ -1 receptor. ( C ) Detection of ASIC1a in a membrane fraction from stably transfected cells (M) and after isolation (I) by binding to Ni 2+ -agarose. Samples were analyzed by SDS-PAGE and either silver staining ( left panel ) or immunoblotting ( right panel ) using rabbit polyclonal anti-ASIC1a antibody, followed by a horseradish peroxidase-conjugated goat anti-rabbit secondary antibody. Immunoreactive bands were visualized using enhanced chemiluminescence. Arrowheads indicate molecular mass markers (kDa). ( D ) Detection of ASIC1a and the σ -1 receptor after isolation from σ -1 receptor-transfected cells by binding to Ni 2+ -agarose. Samples were analyzed by SDS-PAGE and either silver staining ( left panel ) or immunoblotting ( right panel ) using anti-ASIC1a antibody or mouse monoclonal anti-FLAG antibody. ( E ) A screen for the presence of endogenous σ -1 receptor in protein samples isolated from nontransfected ASIC1a-expressing cells or σ -1 receptor-transfected cells. Samples were analyzed by SDS-PAGE and immunoblotting using anti-ASIC1a antibody, anti-FLAG antibody, or rabbit polyclonal anti- σ -1 receptor antibody.
Techniques Used: Isolation, Expressing, Immunofluorescence, Stable Transfection, Transfection, Incubation, Confocal Laser Scanning Microscopy, Binding Assay, SDS Page, Silver Staining

Figure Legend Snippet: Determination of the raft association of ASIC1a and the σ -1 receptor. Cells were extracted with ice-cold Triton X-100 (1%) and subjected to centrifugation on Optiprep density gradients. Fractions from the gradient were analyzed by SDS-PAGE and immunoblotting using anti-ASIC1a antibody, anti-FLAG antibody (for the σ -1 receptor), or anti-caveolin antibody (as a raft marker).
Techniques Used: Centrifugation, SDS Page, Marker