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rdi research diagnostics atpase
Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, <t>OmpA</t> and <t>ATPase</t> were used to monitor fractionation of the outer membrane and inner membranes, respectively.
Atpase, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Green Fluorescent Protein Functions as a Reporter for Protein Localization in Escherichia coli †"

Article Title: Green Fluorescent Protein Functions as a Reporter for Protein Localization in Escherichia coli †

Journal: Journal of Bacteriology

doi:

Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, OmpA and ATPase were used to monitor fractionation of the outer membrane and inner membranes, respectively.
Figure Legend Snippet: Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, OmpA and ATPase were used to monitor fractionation of the outer membrane and inner membranes, respectively.

Techniques Used: Cell Fractionation, Expressing, Molecular Weight, Fractionation

Related Articles

Fractionation:

Article Title: Green Fluorescent Protein Functions as a Reporter for Protein Localization in Escherichia coli †
Article Snippet: .. Antibodies to OmpA and ATPase (Research Diagnostics, Flanders, N.J.) were used to monitor fractionation of the outer and inner membranes, respectively. .. Transformants were cultured in E minimal medium supplemented with amino acids excluding methionine and cysteine to an OD600 of 0.3.

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    rdi research diagnostics α3 na k atpase
    Agrin binds specifically to the <t>α3</t> <t>Na,K-ATPase</t> in heart. A and A ′, low power photomicrographs of a frozen section through an E18 mouse heart ventricle double labeled for agrin ( A ) and α3 Na,K-ATPase ( A ′). Both agrin and the α3 Na,K-ATPase are expressed throughout the ventricular myocardium. Higher levels of α3 Na,K-ATPase can be seen in a subpopulation of cells, possibly developing Purkinje fibers, concentrated in the ventricular septum ( arrows ) and scattered throughout the myocardium. B and B ′, agrin ( B ) and α3 Na,K-ATPase ( B ′) are also expressed in adult cardiac muscle fibers. Viewed at higher magnification, agrin appears relatively evenly distributed, whereas α3 Na,K-ATPase is characterized by the presence of more intensely labeled puncta set against a low background. C , typical Western blots (50 μg/protein/lane) of embryo and adult ventricular muscle probed with antibodies against the α3, α2, or α1 subunit of the Na,K-ATPase. The α3 subunit migrates as a single band of a 110 kDa in saline ( S )-treated control tissue and is more abundant in embryos than in adults. Cross-linking saline-treated tissue with BS 3 generates a ≥300-kDa agrin-α3 subunit complex in which formation is blocked by the presence of either C-Ag15 or C-Ag20, resulting instead in 125- and 130-kDa bands, respectively. In contrast, Western blots of aliquots of the BS 3 cross-linked tissue probed with antibodies to α2 or α1 contain only a single band of 110 kDa, evidence that agrin binds specifically to the α3 Na,K-ATPase. Whereas the level of α2 subunit expression increases during development, the α1 subunit is unchanged, confirming similar loading across lanes. D , tissue samples, treated as in C , were solubilized in detergent-containing buffer and immunoprecipitated ( IP ) with either the anti-agrin serum ( Ag ) or anti-α3 subunit monoclonal antibody (α 3 ), and the immunoprecipitates were analyzed by Western blotting with the anti-agrin serum. Native agrin glycoprotein immunoprecipitated by the anti-agrin serum from control tissue treated with saline alone is shown for comparison and appears as a broad band of ≥200 kDa. Consistent with interaction between endogenous agrin and the α3 subunit, cross-linking results in the appearance of a high molecular mass species of ≥300 kDa recognized by both the anti-α3 and anti-agrin antibodies. Formation of the agrin-α3 complex is blocked by cross-linking in the presence of a saturating concentration of either C-Ag15 or C-Ag20.
    α3 Na K Atpase, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    rdi research diagnostics atpase
    Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, <t>OmpA</t> and <t>ATPase</t> were used to monitor fractionation of the outer membrane and inner membranes, respectively.
    Atpase, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 89/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/atpase/product/rdi research diagnostics
    Average 89 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    atpase - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    85
    rdi research diagnostics rat na k atpase β subunit
    (A) RT-PCR products obtained after 30, 34, or 38 cycles of amplification with primers JR282 and JR286, showing relative expression of Gng3 in various tissues (top three panels), or products from 24 cycles with primers for glyceraldehyde-3-phosphate dehydrogenase ( Gapd ), confirming approximately equal amounts of cDNA in all reactions (bottom panel). (B) (Top) RT-PCR products obtained after 30 cycles of amplification with primers JR282 and JR286 for Gng3 , from cDNA prepared from whole brains of three wild-type, three Gng3 +/− , and three Gng3 −/− mice, without added reverse transcriptase (−RT), or without added cDNA (−cDNA). (Bottom) RT-PCR products obtained after 29 cycles of amplification with primers JR174 and JR175 for elongation factor 1 α2 ( Eef1a2 ), from identical aliquots of the cDNAs. (C) Western blot analysis of proteins prepared from Sf9 cells expressing γ 3 (Std) or brains of three Gng3 −/− , three Gng3 +/− , and three wild-type mice. Top panel shows that γ 3 is reduced in membranes from Gng3 +/− mice and absent in membranes from Gng3 −/− mice. Bottom panel shows that sodium/potassium <t>ATPase</t> <t>β-subunit</t> levels are equal in all lanes.
    Rat Na K Atpase β Subunit, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Agrin binds specifically to the α3 Na,K-ATPase in heart. A and A ′, low power photomicrographs of a frozen section through an E18 mouse heart ventricle double labeled for agrin ( A ) and α3 Na,K-ATPase ( A ′). Both agrin and the α3 Na,K-ATPase are expressed throughout the ventricular myocardium. Higher levels of α3 Na,K-ATPase can be seen in a subpopulation of cells, possibly developing Purkinje fibers, concentrated in the ventricular septum ( arrows ) and scattered throughout the myocardium. B and B ′, agrin ( B ) and α3 Na,K-ATPase ( B ′) are also expressed in adult cardiac muscle fibers. Viewed at higher magnification, agrin appears relatively evenly distributed, whereas α3 Na,K-ATPase is characterized by the presence of more intensely labeled puncta set against a low background. C , typical Western blots (50 μg/protein/lane) of embryo and adult ventricular muscle probed with antibodies against the α3, α2, or α1 subunit of the Na,K-ATPase. The α3 subunit migrates as a single band of a 110 kDa in saline ( S )-treated control tissue and is more abundant in embryos than in adults. Cross-linking saline-treated tissue with BS 3 generates a ≥300-kDa agrin-α3 subunit complex in which formation is blocked by the presence of either C-Ag15 or C-Ag20, resulting instead in 125- and 130-kDa bands, respectively. In contrast, Western blots of aliquots of the BS 3 cross-linked tissue probed with antibodies to α2 or α1 contain only a single band of 110 kDa, evidence that agrin binds specifically to the α3 Na,K-ATPase. Whereas the level of α2 subunit expression increases during development, the α1 subunit is unchanged, confirming similar loading across lanes. D , tissue samples, treated as in C , were solubilized in detergent-containing buffer and immunoprecipitated ( IP ) with either the anti-agrin serum ( Ag ) or anti-α3 subunit monoclonal antibody (α 3 ), and the immunoprecipitates were analyzed by Western blotting with the anti-agrin serum. Native agrin glycoprotein immunoprecipitated by the anti-agrin serum from control tissue treated with saline alone is shown for comparison and appears as a broad band of ≥200 kDa. Consistent with interaction between endogenous agrin and the α3 subunit, cross-linking results in the appearance of a high molecular mass species of ≥300 kDa recognized by both the anti-α3 and anti-agrin antibodies. Formation of the agrin-α3 complex is blocked by cross-linking in the presence of a saturating concentration of either C-Ag15 or C-Ag20.

    Journal: The Journal of Biological Chemistry

    Article Title: Agrin Regulation of ?3 Sodium-Potassium ATPase Activity Modulates Cardiac Myocyte Contraction *

    doi: 10.1074/jbc.M806855200

    Figure Lengend Snippet: Agrin binds specifically to the α3 Na,K-ATPase in heart. A and A ′, low power photomicrographs of a frozen section through an E18 mouse heart ventricle double labeled for agrin ( A ) and α3 Na,K-ATPase ( A ′). Both agrin and the α3 Na,K-ATPase are expressed throughout the ventricular myocardium. Higher levels of α3 Na,K-ATPase can be seen in a subpopulation of cells, possibly developing Purkinje fibers, concentrated in the ventricular septum ( arrows ) and scattered throughout the myocardium. B and B ′, agrin ( B ) and α3 Na,K-ATPase ( B ′) are also expressed in adult cardiac muscle fibers. Viewed at higher magnification, agrin appears relatively evenly distributed, whereas α3 Na,K-ATPase is characterized by the presence of more intensely labeled puncta set against a low background. C , typical Western blots (50 μg/protein/lane) of embryo and adult ventricular muscle probed with antibodies against the α3, α2, or α1 subunit of the Na,K-ATPase. The α3 subunit migrates as a single band of a 110 kDa in saline ( S )-treated control tissue and is more abundant in embryos than in adults. Cross-linking saline-treated tissue with BS 3 generates a ≥300-kDa agrin-α3 subunit complex in which formation is blocked by the presence of either C-Ag15 or C-Ag20, resulting instead in 125- and 130-kDa bands, respectively. In contrast, Western blots of aliquots of the BS 3 cross-linked tissue probed with antibodies to α2 or α1 contain only a single band of 110 kDa, evidence that agrin binds specifically to the α3 Na,K-ATPase. Whereas the level of α2 subunit expression increases during development, the α1 subunit is unchanged, confirming similar loading across lanes. D , tissue samples, treated as in C , were solubilized in detergent-containing buffer and immunoprecipitated ( IP ) with either the anti-agrin serum ( Ag ) or anti-α3 subunit monoclonal antibody (α 3 ), and the immunoprecipitates were analyzed by Western blotting with the anti-agrin serum. Native agrin glycoprotein immunoprecipitated by the anti-agrin serum from control tissue treated with saline alone is shown for comparison and appears as a broad band of ≥200 kDa. Consistent with interaction between endogenous agrin and the α3 subunit, cross-linking results in the appearance of a high molecular mass species of ≥300 kDa recognized by both the anti-α3 and anti-agrin antibodies. Formation of the agrin-α3 complex is blocked by cross-linking in the presence of a saturating concentration of either C-Ag15 or C-Ag20.

    Article Snippet: Mouse monoclonal antibodies against the α3 Na,K-ATPase (XVIF9-G10, Research Diagnostics, Inc. ( )) and α1 Na,K-ATPase (clone 9A-5; A275, Sigma-Aldrich ( )) and rabbit serum against the α2 Na,K-ATPase (AB9094, Millipore ( , )) were used at a dilution of 1:200.

    Techniques: Labeling, Western Blot, Expressing, Immunoprecipitation, Concentration Assay

    Agrin modulates cytoplasmic Na + in cultured cardiac myocytes. A , record from a single cultured cardiac myocyte showing small, spontaneous fluctuations in cytoplasmic Na + observed in normal saline ( S ) and larger, sustained increase in Na + following treatment with agrin (C-Ag20). B , bars show mean change in Na + concentration in response to saturating concentration of C-Ag20 or ouabain ( Oaub ). To control for differences in SBFI loading between cells and experiments, data for each cell was base line-subtracted and normalized to the maximal response to a mixture of ouabain and gramicidin. Treatment with agrin results in a significant increase in Na + levels, albeit lower than that observed in the presence of ouabain, a pan-specific Na,K-ATPase inhibitor. C , whereas treatment with C-Ag15 alone had no effect on Na + , C-Ag15 clearly antagonized the response to C-Ag20. Note that the concentration of C-Ag20 used for the competition assays was 10-fold lower than in B , resulting in a smaller maximal response to C-Ag20. Bars show data from 14–27 cells from a minimum of two experiments. **, p

    Journal: The Journal of Biological Chemistry

    Article Title: Agrin Regulation of ?3 Sodium-Potassium ATPase Activity Modulates Cardiac Myocyte Contraction *

    doi: 10.1074/jbc.M806855200

    Figure Lengend Snippet: Agrin modulates cytoplasmic Na + in cultured cardiac myocytes. A , record from a single cultured cardiac myocyte showing small, spontaneous fluctuations in cytoplasmic Na + observed in normal saline ( S ) and larger, sustained increase in Na + following treatment with agrin (C-Ag20). B , bars show mean change in Na + concentration in response to saturating concentration of C-Ag20 or ouabain ( Oaub ). To control for differences in SBFI loading between cells and experiments, data for each cell was base line-subtracted and normalized to the maximal response to a mixture of ouabain and gramicidin. Treatment with agrin results in a significant increase in Na + levels, albeit lower than that observed in the presence of ouabain, a pan-specific Na,K-ATPase inhibitor. C , whereas treatment with C-Ag15 alone had no effect on Na + , C-Ag15 clearly antagonized the response to C-Ag20. Note that the concentration of C-Ag20 used for the competition assays was 10-fold lower than in B , resulting in a smaller maximal response to C-Ag20. Bars show data from 14–27 cells from a minimum of two experiments. **, p

    Article Snippet: Mouse monoclonal antibodies against the α3 Na,K-ATPase (XVIF9-G10, Research Diagnostics, Inc. ( )) and α1 Na,K-ATPase (clone 9A-5; A275, Sigma-Aldrich ( )) and rabbit serum against the α2 Na,K-ATPase (AB9094, Millipore ( , )) were used at a dilution of 1:200.

    Techniques: Cell Culture, Concentration Assay

    Agrin-dependent tyrosine phosphorylation of the α3 subunit of the Na,K-ATPase. A , typical Western blots show α1, α2, and α3 Na,K-ATPase subunits immunoprecipitated from adult ventricular muscle by an anti-phosphotyrosine antibody. C-Ag15 reduced basal levels of phosphorylation only in the α3 subunit. Treatment with C-Ag20 increased the level of α3 subunit phosphorylation but had no effect on either α1 or α2. Endogenous phosphorylation of all three subunits and C-Ag20-induced phosphorylation of the α3 subunit is blocked by genistein ( Gen ). B , densitometric analysis of five independent experiments similar to that shown in A . Tyrosine phosphorylation ( PY ) of the α3 subunit is decreased following treatment with C-Ag15 but increased in the presence of C-Ag20. C-Ag20-dependent phosphorylation of the α3 subunit was blocked by genistein, suggesting that agrin interaction with the α3 Na,K-ATPase activates a tyrosine kinase. **, p

    Journal: The Journal of Biological Chemistry

    Article Title: Agrin Regulation of ?3 Sodium-Potassium ATPase Activity Modulates Cardiac Myocyte Contraction *

    doi: 10.1074/jbc.M806855200

    Figure Lengend Snippet: Agrin-dependent tyrosine phosphorylation of the α3 subunit of the Na,K-ATPase. A , typical Western blots show α1, α2, and α3 Na,K-ATPase subunits immunoprecipitated from adult ventricular muscle by an anti-phosphotyrosine antibody. C-Ag15 reduced basal levels of phosphorylation only in the α3 subunit. Treatment with C-Ag20 increased the level of α3 subunit phosphorylation but had no effect on either α1 or α2. Endogenous phosphorylation of all three subunits and C-Ag20-induced phosphorylation of the α3 subunit is blocked by genistein ( Gen ). B , densitometric analysis of five independent experiments similar to that shown in A . Tyrosine phosphorylation ( PY ) of the α3 subunit is decreased following treatment with C-Ag15 but increased in the presence of C-Ag20. C-Ag20-dependent phosphorylation of the α3 subunit was blocked by genistein, suggesting that agrin interaction with the α3 Na,K-ATPase activates a tyrosine kinase. **, p

    Article Snippet: Mouse monoclonal antibodies against the α3 Na,K-ATPase (XVIF9-G10, Research Diagnostics, Inc. ( )) and α1 Na,K-ATPase (clone 9A-5; A275, Sigma-Aldrich ( )) and rabbit serum against the α2 Na,K-ATPase (AB9094, Millipore ( , )) were used at a dilution of 1:200.

    Techniques: Western Blot, Immunoprecipitation

    Agrin regulates α3 Na,K-ATPase activity. A , the production of inorganic phosphate from ATP by purified P0 ventricular myocyte sarcolemmal membranes was measured, and background ATPase activity, defined as the ouabain-insensitive component, was subtracted. Na,K-ATPase activity in control, untreated myocyte membranes ( filled circle ) is shown for reference. Measurement of ATP hydrolysis in the presence of different concentrations of the agrin fragments shows that C-Ag20 significantly inhibits ( p

    Journal: The Journal of Biological Chemistry

    Article Title: Agrin Regulation of ?3 Sodium-Potassium ATPase Activity Modulates Cardiac Myocyte Contraction *

    doi: 10.1074/jbc.M806855200

    Figure Lengend Snippet: Agrin regulates α3 Na,K-ATPase activity. A , the production of inorganic phosphate from ATP by purified P0 ventricular myocyte sarcolemmal membranes was measured, and background ATPase activity, defined as the ouabain-insensitive component, was subtracted. Na,K-ATPase activity in control, untreated myocyte membranes ( filled circle ) is shown for reference. Measurement of ATP hydrolysis in the presence of different concentrations of the agrin fragments shows that C-Ag20 significantly inhibits ( p

    Article Snippet: Mouse monoclonal antibodies against the α3 Na,K-ATPase (XVIF9-G10, Research Diagnostics, Inc. ( )) and α1 Na,K-ATPase (clone 9A-5; A275, Sigma-Aldrich ( )) and rabbit serum against the α2 Na,K-ATPase (AB9094, Millipore ( , )) were used at a dilution of 1:200.

    Techniques: Activity Assay, Purification

    Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, OmpA and ATPase were used to monitor fractionation of the outer membrane and inner membranes, respectively.

    Journal: Journal of Bacteriology

    Article Title: Green Fluorescent Protein Functions as a Reporter for Protein Localization in Escherichia coli †

    doi:

    Figure Lengend Snippet: Cell fractionation of shortened MBP-GFP hybrid protein. Cells expressing MBP[SS128]-GFP were fractionated into inner and outer membranes as described in Materials and Methods. The immunoblot shown was decorated with anti-GFP antibody. MBP[SS128]-GFP migrated at a predicted molecular mass of 40 kDa and fractionated with the outer membrane. An apparent smaller-molecular-weight derivative of this protein was also found in the inner membrane fraction. Molecular weight (MW) standards are as shown. IM, inner membrane fraction; IM-W, KCl wash of inner membrane fraction; OM, outer membrane fraction; OM-W, KCl wash of outer membrane fraction. Although not shown, OmpA and ATPase were used to monitor fractionation of the outer membrane and inner membranes, respectively.

    Article Snippet: Antibodies to OmpA and ATPase (Research Diagnostics, Flanders, N.J.) were used to monitor fractionation of the outer and inner membranes, respectively.

    Techniques: Cell Fractionation, Expressing, Molecular Weight, Fractionation

    Expression of CCK2R in normal corpus A–C , CCK2R immunoreactivity in section of full-thickness gastric corpus. A , 4′,6-diamidino-2-phenylindole (DAPI); B , CCK2R (fluorescein isothiocyanate; FITC); and C , overlay. D–G , CCK2R and H + ,K + -ATPase β subunit immunoreactivity in gastric corpus. D , DAPI; E , CCK2R (FITC); F , H + ,K + -ATPase β subunit (Texas Red); and G , CCK2R and H + ,K + -ATPase β subunit overlay. H–K , CCK2R and VMAT2 immunoreactivity in gastric corpus. H , DAPI; I , CCK2R (FITC); J , VMAT2 (Texas Red); and K , CCK2R and VMAT2 overlay. Sections are representative of samples obtained from four individual animals. For A–C , scale bar respresents 250 μm; for D–K , scale bar represents 200 μm.

    Journal: Experimental Physiology

    Article Title: Regulation of mammalian gastrin/CCK receptor (CCK2R) expression in vitro and in vivo

    doi: 10.1113/expphysiol.2007.040683

    Figure Lengend Snippet: Expression of CCK2R in normal corpus A–C , CCK2R immunoreactivity in section of full-thickness gastric corpus. A , 4′,6-diamidino-2-phenylindole (DAPI); B , CCK2R (fluorescein isothiocyanate; FITC); and C , overlay. D–G , CCK2R and H + ,K + -ATPase β subunit immunoreactivity in gastric corpus. D , DAPI; E , CCK2R (FITC); F , H + ,K + -ATPase β subunit (Texas Red); and G , CCK2R and H + ,K + -ATPase β subunit overlay. H–K , CCK2R and VMAT2 immunoreactivity in gastric corpus. H , DAPI; I , CCK2R (FITC); J , VMAT2 (Texas Red); and K , CCK2R and VMAT2 overlay. Sections are representative of samples obtained from four individual animals. For A–C , scale bar respresents 250 μm; for D–K , scale bar represents 200 μm.

    Article Snippet: Antibodies to vesicular monoamine transporter 2 (VMAT2), the gastric H+ ,K+ -ATPase β-subunit, desmin, smooth muscle α-actin and vimentin were from Research Diagnostics (Flanders, NJ, USA).

    Techniques: Expressing

    (A) RT-PCR products obtained after 30, 34, or 38 cycles of amplification with primers JR282 and JR286, showing relative expression of Gng3 in various tissues (top three panels), or products from 24 cycles with primers for glyceraldehyde-3-phosphate dehydrogenase ( Gapd ), confirming approximately equal amounts of cDNA in all reactions (bottom panel). (B) (Top) RT-PCR products obtained after 30 cycles of amplification with primers JR282 and JR286 for Gng3 , from cDNA prepared from whole brains of three wild-type, three Gng3 +/− , and three Gng3 −/− mice, without added reverse transcriptase (−RT), or without added cDNA (−cDNA). (Bottom) RT-PCR products obtained after 29 cycles of amplification with primers JR174 and JR175 for elongation factor 1 α2 ( Eef1a2 ), from identical aliquots of the cDNAs. (C) Western blot analysis of proteins prepared from Sf9 cells expressing γ 3 (Std) or brains of three Gng3 −/− , three Gng3 +/− , and three wild-type mice. Top panel shows that γ 3 is reduced in membranes from Gng3 +/− mice and absent in membranes from Gng3 −/− mice. Bottom panel shows that sodium/potassium ATPase β-subunit levels are equal in all lanes.

    Journal: Molecular and Cellular Biology

    Article Title: Mice with Deficiency of G Protein ?3 Are Lean and Have Seizures

    doi: 10.1128/MCB.24.17.7758-7768.2004

    Figure Lengend Snippet: (A) RT-PCR products obtained after 30, 34, or 38 cycles of amplification with primers JR282 and JR286, showing relative expression of Gng3 in various tissues (top three panels), or products from 24 cycles with primers for glyceraldehyde-3-phosphate dehydrogenase ( Gapd ), confirming approximately equal amounts of cDNA in all reactions (bottom panel). (B) (Top) RT-PCR products obtained after 30 cycles of amplification with primers JR282 and JR286 for Gng3 , from cDNA prepared from whole brains of three wild-type, three Gng3 +/− , and three Gng3 −/− mice, without added reverse transcriptase (−RT), or without added cDNA (−cDNA). (Bottom) RT-PCR products obtained after 29 cycles of amplification with primers JR174 and JR175 for elongation factor 1 α2 ( Eef1a2 ), from identical aliquots of the cDNAs. (C) Western blot analysis of proteins prepared from Sf9 cells expressing γ 3 (Std) or brains of three Gng3 −/− , three Gng3 +/− , and three wild-type mice. Top panel shows that γ 3 is reduced in membranes from Gng3 +/− mice and absent in membranes from Gng3 −/− mice. Bottom panel shows that sodium/potassium ATPase β-subunit levels are equal in all lanes.

    Article Snippet: Antisera for Ras (BD Biosciences) and for the rat Na+ /K+ -ATPase β subunit (Research Diagnostics, Inc., Flanders, N.J.) were used at a 1:2,000 dilution.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Expressing, Mouse Assay, Western Blot