Structured Review

Becton Dickinson na k atpase β2 subunit
<t>Na,K-ATPase</t> β 2 and α 2 subunits are localized almost exclusively in T-tubules in cardiomyocytes, whereas the α 1 and β 1 subunits are localized in both sarcolemma and T-tubules. A , frozen sections of rat heart were double-stained
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Images

1) Product Images from "Selective Assembly of Na,K-ATPase α2β2 Heterodimers in the Heart"

Article Title: Selective Assembly of Na,K-ATPase α2β2 Heterodimers in the Heart

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M116.751735

Na,K-ATPase β 2 and α 2 subunits are localized almost exclusively in T-tubules in cardiomyocytes, whereas the α 1 and β 1 subunits are localized in both sarcolemma and T-tubules. A , frozen sections of rat heart were double-stained
Figure Legend Snippet: Na,K-ATPase β 2 and α 2 subunits are localized almost exclusively in T-tubules in cardiomyocytes, whereas the α 1 and β 1 subunits are localized in both sarcolemma and T-tubules. A , frozen sections of rat heart were double-stained

Techniques Used: Staining

Expression of purified Na,K-ATPase isoforms. Coomassie-stained SDS-PAGE of purified isoforms (5 μg/lane). For deglycosylation, samples were denatured and treated with PNGase F for 60 min at 37 °C.
Figure Legend Snippet: Expression of purified Na,K-ATPase isoforms. Coomassie-stained SDS-PAGE of purified isoforms (5 μg/lane). For deglycosylation, samples were denatured and treated with PNGase F for 60 min at 37 °C.

Techniques Used: Expressing, Purification, Staining, SDS Page

Models for docking of DcB to α 1 β 1 , α 2 β 2 , and α 2 β 3 . DcB was docked into homology models of human Na,K-ATPase isoforms derived from the porcine E2P·Mg·digoxin structure (Protein Data Bank code
Figure Legend Snippet: Models for docking of DcB to α 1 β 1 , α 2 β 2 , and α 2 β 3 . DcB was docked into homology models of human Na,K-ATPase isoforms derived from the porcine E2P·Mg·digoxin structure (Protein Data Bank code

Techniques Used: Derivative Assay

The Na,K-ATPase α 2 subunit and β 2 subunit preferentially interact with each other in mouse heart. A , Western blotting analysis of proteins immunoprecipitated and co-immunoprecipitated from the detergent extracts of mouse heart microsome
Figure Legend Snippet: The Na,K-ATPase α 2 subunit and β 2 subunit preferentially interact with each other in mouse heart. A , Western blotting analysis of proteins immunoprecipitated and co-immunoprecipitated from the detergent extracts of mouse heart microsome

Techniques Used: Western Blot, Immunoprecipitation

The Na,K-ATPase α 2 subunit and β 2 subunit are preferentially expressed in the mouse embryonic heart in contrast to the ubiquitously expressed α 1 subunit. A , paraffin-embedded sections of mouse embryos (embryonic day 12.5) were
Figure Legend Snippet: The Na,K-ATPase α 2 subunit and β 2 subunit are preferentially expressed in the mouse embryonic heart in contrast to the ubiquitously expressed α 1 subunit. A , paraffin-embedded sections of mouse embryos (embryonic day 12.5) were

Techniques Used:

Potassium-digoxin antagonism depicted for α 1 β 1 , α 2 β 1 , α 2 β 2 , and α 2 β 3 . Inhibition of purified Na,K-ATPase isoforms by digoxin was measured, and K i values were plotted against the potassium
Figure Legend Snippet: Potassium-digoxin antagonism depicted for α 1 β 1 , α 2 β 1 , α 2 β 2 , and α 2 β 3 . Inhibition of purified Na,K-ATPase isoforms by digoxin was measured, and K i values were plotted against the potassium

Techniques Used: Inhibition, Purification

Potassium-[ 3 H]digoxin antagonism. Yeast membranes harboring α 2 β 1 , α 2 β 2 , or α 2 β 3 Na,K-ATPase were incubated with varying concentrations of KCl, and residual [ 3 H]digoxin binding was measured. The lines represent
Figure Legend Snippet: Potassium-[ 3 H]digoxin antagonism. Yeast membranes harboring α 2 β 1 , α 2 β 2 , or α 2 β 3 Na,K-ATPase were incubated with varying concentrations of KCl, and residual [ 3 H]digoxin binding was measured. The lines represent

Techniques Used: Incubation, Binding Assay

Inhibition of Na,K-ATPase Isoforms by Perhydro-1,4-oxazepine Derivatives of Digoxin
Figure Legend Snippet: Inhibition of Na,K-ATPase Isoforms by Perhydro-1,4-oxazepine Derivatives of Digoxin

Techniques Used: Inhibition

2) Product Images from "Subunit Isoform Selectivity in Assembly of Na,K-ATPase ?-? Heterodimers *"

Article Title: Subunit Isoform Selectivity in Assembly of Na,K-ATPase ?-? Heterodimers *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M112.370734

Localization of the Na,K-ATPase β 2 subunit in nerve sections is similar to that of the Na,K-ATPase α 2 subunit, but not of the Na,K-ATPase α 1 subunit or β 1 subunit. A , frozen sections of rat sciatic nerve were double stained
Figure Legend Snippet: Localization of the Na,K-ATPase β 2 subunit in nerve sections is similar to that of the Na,K-ATPase α 2 subunit, but not of the Na,K-ATPase α 1 subunit or β 1 subunit. A , frozen sections of rat sciatic nerve were double stained

Techniques Used: Staining

The Na,K-ATPase α 2 -β 2 complex is less stable than the Na,K-ATPase α-β 1 or α-β 3 complexes in detergent extracts obtained from mouse brain membranes. A , various concentrations of DDM were used to extract proteins
Figure Legend Snippet: The Na,K-ATPase α 2 -β 2 complex is less stable than the Na,K-ATPase α-β 1 or α-β 3 complexes in detergent extracts obtained from mouse brain membranes. A , various concentrations of DDM were used to extract proteins

Techniques Used:

Localization of the Na,K-ATPase α 1 , α 2 , and β 2 subunits in rat kidney and brain sections. Frozen sections of rat kidney ( A ) and rat brain ( B ) were double-stained by using mouse antibodies against α 1 subunit ( green ) and
Figure Legend Snippet: Localization of the Na,K-ATPase α 1 , α 2 , and β 2 subunits in rat kidney and brain sections. Frozen sections of rat kidney ( A ) and rat brain ( B ) were double-stained by using mouse antibodies against α 1 subunit ( green ) and

Techniques Used: Staining

The Na,K-ATPase α 1 -β 1 complex is more stable than the Na,K-ATPase α 1 -β 2 complex in detergent extracts from MDCK cells. A , MDCK cells stably expressing either YFP-β 1 or YFP-β 2 were lysed by incubation with
Figure Legend Snippet: The Na,K-ATPase α 1 -β 1 complex is more stable than the Na,K-ATPase α 1 -β 2 complex in detergent extracts from MDCK cells. A , MDCK cells stably expressing either YFP-β 1 or YFP-β 2 were lysed by incubation with

Techniques Used: Stable Transfection, Expressing, Incubation

The Na,K-ATPase α 1 -β 2 complex is preserved in digitonin and CHAPS extracts from MDCK cells. A , proteins were extracted from MDCK cells expressing YFP-β 1 , or YFP-β 2 , or YFP-linked bile acid transporter (YFP-NTCP), with the
Figure Legend Snippet: The Na,K-ATPase α 1 -β 2 complex is preserved in digitonin and CHAPS extracts from MDCK cells. A , proteins were extracted from MDCK cells expressing YFP-β 1 , or YFP-β 2 , or YFP-linked bile acid transporter (YFP-NTCP), with the

Techniques Used: Expressing

The Na,K-ATPase α 2 and β 2 subunits are selectively co-immunoprecipitated from mouse brain extracts. Proteins were extracted from mouse brain homogenate by using 1% Nonidet P-40/0.5% DOC. A , the antibodies against the Na,K-ATPase β
Figure Legend Snippet: The Na,K-ATPase α 2 and β 2 subunits are selectively co-immunoprecipitated from mouse brain extracts. Proteins were extracted from mouse brain homogenate by using 1% Nonidet P-40/0.5% DOC. A , the antibodies against the Na,K-ATPase β

Techniques Used: Immunoprecipitation

Related Articles

Staining:

Article Title: Subunit Isoform Selectivity in Assembly of Na,K-ATPase ?-? Heterodimers *
Article Snippet: Paragraph title: Primary Antibodies Used for Immunofluorescent Staining and Western Blot Analysis ... Also, the following monoclonal antibodies were used for Western blot analysis: against GFP, clones 7.1 and 13.1, which also recognizes YFP (Roche Diagnostics), against the Na,K-ATPase α1 subunit, clone C464.6 (Millipore), against the Na,K-ATPase α3 subunit (Upstate), against the Na,K-ATPase β2 subunit, clone 35 (BD Bioscience Pharmingen), and against the Na,K-ATPase β3 subunit (Santa Cruz Biotechnology).

Article Title: Selective Assembly of Na,K-ATPase α2β2 Heterodimers in the Heart
Article Snippet: For immunofluorescent staining, the following monoclonal antibodies were used: Na,K-ATPase α1 subunit (mouse, clone C464.6, 1:20; Millipore) and Na,K-ATPase β1 subunit (mouse, clone M17 P5 F11, 1:100; Affinity Bioreagents). .. For Western blotting analysis, the following monoclonal antibodies were used: against the Na,K-ATPase α1 subunit (mouse, clone C464.6, 1:1000; Millipore), against the Na,K-ATPase β2 subunit (mouse, clone 35; BD Transduction Laboratories), and Na,K-ATPase β3 subunit (goat, 1:500; Santa Cruz Biotechnology, Inc.).

Incubation:

Article Title: Akt-Dependent Cell Size Regulation by the Adhesion Molecule on Glia Occurs Independently of Phosphatidylinositol 3-Kinase and Rheb Signaling
Article Snippet: Membranes were then incubated with primary antibodies in TBS-T with 5% nonfat dry milk or 5% bovine serum albumin (for phospho antibodies), washed, and incubated with the appropriate secondary antibody in TBS-T with 5% nonfat dry milk. .. The following commercial primary antibodies were used: anti-PDK1 (3062), anti-Akt (9272), anti-S6K (9202), anti-S6 (2212), anti-MAPK (9102), anti-Rheb (4935), anti-P-S6K (9205), anti-P-S6 (2215), anti-P-MAPK (9101), anti-P-Akt (9271), anti-Gab1 (3232), anti-Na+ /K+ ATPase β2 subunit (AMOG; 610914; BD Transduction Laboratories), anti-Na+ /K+ ATPase β3 subunit ( ; BD Transduction Laboratories), anti-Myc (sc-40; Santa Cruz Biotechnology, Santa Cruz, Calif.), anti-cleaved caspase 3 (9661), anti-P-src (2101), anti-P-insulin-like growth factor receptor (IGFR) (Tyr1131/1426; 3021), anti-P-Tyr (PY20; BD Transduction Laboratories), anti-fibroblast growth factor receptor-1 (FGFR-1; (34720), anti-epidermal growth factor receptor (EGFR) (2232), anti-platelet-derived growth factor receptor (PDGFR) (3162), anti-P-tuberin Ser939, Thr1462, and Tyr1571 (3611, 3614, 3615), anti-P-tyr p85 PI3K binding motif (3821), and anti-α-tubulin (T-9026; Sigma).

Binding Assay:

Article Title: Akt-Dependent Cell Size Regulation by the Adhesion Molecule on Glia Occurs Independently of Phosphatidylinositol 3-Kinase and Rheb Signaling
Article Snippet: .. The following commercial primary antibodies were used: anti-PDK1 (3062), anti-Akt (9272), anti-S6K (9202), anti-S6 (2212), anti-MAPK (9102), anti-Rheb (4935), anti-P-S6K (9205), anti-P-S6 (2215), anti-P-MAPK (9101), anti-P-Akt (9271), anti-Gab1 (3232), anti-Na+ /K+ ATPase β2 subunit (AMOG; 610914; BD Transduction Laboratories), anti-Na+ /K+ ATPase β3 subunit ( ; BD Transduction Laboratories), anti-Myc (sc-40; Santa Cruz Biotechnology, Santa Cruz, Calif.), anti-cleaved caspase 3 (9661), anti-P-src (2101), anti-P-insulin-like growth factor receptor (IGFR) (Tyr1131/1426; 3021), anti-P-Tyr (PY20; BD Transduction Laboratories), anti-fibroblast growth factor receptor-1 (FGFR-1; (34720), anti-epidermal growth factor receptor (EGFR) (2232), anti-platelet-derived growth factor receptor (PDGFR) (3162), anti-P-tuberin Ser939, Thr1462, and Tyr1571 (3611, 3614, 3615), anti-P-tyr p85 PI3K binding motif (3821), and anti-α-tubulin (T-9026; Sigma). ..

Clone Assay:

Article Title: Subunit Isoform Selectivity in Assembly of Na,K-ATPase ?-? Heterodimers *
Article Snippet: .. Also, the following monoclonal antibodies were used for Western blot analysis: against GFP, clones 7.1 and 13.1, which also recognizes YFP (Roche Diagnostics), against the Na,K-ATPase α1 subunit, clone C464.6 (Millipore), against the Na,K-ATPase α3 subunit (Upstate), against the Na,K-ATPase β2 subunit, clone 35 (BD Bioscience Pharmingen), and against the Na,K-ATPase β3 subunit (Santa Cruz Biotechnology). .. Confluent MDCK cell monolayers grown in six-well plates were rinsed twice with ice-cold PBS and incubated with 200 μl/well of the extraction buffer at 4 °C for 30 min followed by scraping cells.

Western Blot:

Article Title: Subunit Isoform Selectivity in Assembly of Na,K-ATPase ?-? Heterodimers *
Article Snippet: .. Also, the following monoclonal antibodies were used for Western blot analysis: against GFP, clones 7.1 and 13.1, which also recognizes YFP (Roche Diagnostics), against the Na,K-ATPase α1 subunit, clone C464.6 (Millipore), against the Na,K-ATPase α3 subunit (Upstate), against the Na,K-ATPase β2 subunit, clone 35 (BD Bioscience Pharmingen), and against the Na,K-ATPase β3 subunit (Santa Cruz Biotechnology). .. Confluent MDCK cell monolayers grown in six-well plates were rinsed twice with ice-cold PBS and incubated with 200 μl/well of the extraction buffer at 4 °C for 30 min followed by scraping cells.

Article Title: Akt-Dependent Cell Size Regulation by the Adhesion Molecule on Glia Occurs Independently of Phosphatidylinositol 3-Kinase and Rheb Signaling
Article Snippet: Paragraph title: Western blotting and antibodies. ... The following commercial primary antibodies were used: anti-PDK1 (3062), anti-Akt (9272), anti-S6K (9202), anti-S6 (2212), anti-MAPK (9102), anti-Rheb (4935), anti-P-S6K (9205), anti-P-S6 (2215), anti-P-MAPK (9101), anti-P-Akt (9271), anti-Gab1 (3232), anti-Na+ /K+ ATPase β2 subunit (AMOG; 610914; BD Transduction Laboratories), anti-Na+ /K+ ATPase β3 subunit ( ; BD Transduction Laboratories), anti-Myc (sc-40; Santa Cruz Biotechnology, Santa Cruz, Calif.), anti-cleaved caspase 3 (9661), anti-P-src (2101), anti-P-insulin-like growth factor receptor (IGFR) (Tyr1131/1426; 3021), anti-P-Tyr (PY20; BD Transduction Laboratories), anti-fibroblast growth factor receptor-1 (FGFR-1; (34720), anti-epidermal growth factor receptor (EGFR) (2232), anti-platelet-derived growth factor receptor (PDGFR) (3162), anti-P-tuberin Ser939, Thr1462, and Tyr1571 (3611, 3614, 3615), anti-P-tyr p85 PI3K binding motif (3821), and anti-α-tubulin (T-9026; Sigma).

Article Title: Selective Assembly of Na,K-ATPase α2β2 Heterodimers in the Heart
Article Snippet: .. For Western blotting analysis, the following monoclonal antibodies were used: against the Na,K-ATPase α1 subunit (mouse, clone C464.6, 1:1000; Millipore), against the Na,K-ATPase β2 subunit (mouse, clone 35; BD Transduction Laboratories), and Na,K-ATPase β3 subunit (goat, 1:500; Santa Cruz Biotechnology, Inc.). .. Na,K-ATPase β1 subunit polyclonal antibody (rabbit; 1:5000) was a generous gift of Dr. W. James Ball, Jr. (University of Cincinnati).