rabbit anti psd93  (Alomone Labs)


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    Alomone Labs rabbit anti psd93
    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for <t>PSD93.</t> B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.
    Rabbit Anti Psd93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti psd93/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti psd93 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation"

    Article Title: Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0023978

    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.
    Figure Legend Snippet: A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.

    Techniques Used: Western Blot, Staining, Lysis, Centrifugation, Binding Assay, Dot Blot

    rabbit anti psd93  (Alomone Labs)


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    Alomone Labs rabbit anti psd93
    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for <t>PSD93.</t> B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.
    Rabbit Anti Psd93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti psd93/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti psd93 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation"

    Article Title: Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0023978

    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.
    Figure Legend Snippet: A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.

    Techniques Used: Western Blot, Staining, Lysis, Centrifugation, Binding Assay, Dot Blot

    anti psd 93  (Alomone Labs)


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    Alomone Labs anti psd 93
    Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: <t>PSD-93,</t> 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.
    Anti Psd 93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti psd 93/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti psd 93 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Impaired synaptic clustering of postsynaptic density proteins and altered signal transmission in hippocampal neurons, and disrupted learning behavior in PDZ1 and PDZ2 ligand binding-deficient PSD-95 knockin mice"

    Article Title: Impaired synaptic clustering of postsynaptic density proteins and altered signal transmission in hippocampal neurons, and disrupted learning behavior in PDZ1 and PDZ2 ligand binding-deficient PSD-95 knockin mice

    Journal: Molecular Brain

    doi: 10.1186/1756-6606-5-43

    Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: PSD-93, 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.
    Figure Legend Snippet: Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: PSD-93, 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.

    Techniques Used: Western Blot, Quantitation Assay, Mutagenesis

    Altered expression of SAP102 and PSD-93 in the hippocampus of KI mice. A , Immunoblot analyses of hippocampal homogenates from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of homogenate proteins were analyzed per lane: PSD-93 and SAP102, WT and KI: 5 μg; GluA1 and GluA2/3, WT and KI: 10 μg; and PSD-95, WT: 5 μg, KI: 20 μg. The arrowhead indicates the degraded bands of PSD-93. B , Quantitation of PSD proteins in the hippocampal homogenates based on the results shown in A , normalized to the wild-type levels at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of hippocampal homogenates at various ages (P4–P65) were examined. The means and SEM of WT and KI data were compared using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD homogenates, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, and increased levels of SAP102 were observed during development compared to the WT mice, while the levels of GluA1 and GluA2/3 were not significantly different from those of the WT mice. C , Northern blot analyses of PSD-95 and SAP102 mRNA. Ten micrograms of total RNA prepared from WT and KI mice hippocampi were analyzed in a formaldehyde-containing 1% agarose gel. The numbers on the left side indicate the sizes of the digoxigenin-labeled RNA molecular weight markers. Arrowheads indicate the major band for each mRNA.
    Figure Legend Snippet: Altered expression of SAP102 and PSD-93 in the hippocampus of KI mice. A , Immunoblot analyses of hippocampal homogenates from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of homogenate proteins were analyzed per lane: PSD-93 and SAP102, WT and KI: 5 μg; GluA1 and GluA2/3, WT and KI: 10 μg; and PSD-95, WT: 5 μg, KI: 20 μg. The arrowhead indicates the degraded bands of PSD-93. B , Quantitation of PSD proteins in the hippocampal homogenates based on the results shown in A , normalized to the wild-type levels at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of hippocampal homogenates at various ages (P4–P65) were examined. The means and SEM of WT and KI data were compared using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD homogenates, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, and increased levels of SAP102 were observed during development compared to the WT mice, while the levels of GluA1 and GluA2/3 were not significantly different from those of the WT mice. C , Northern blot analyses of PSD-95 and SAP102 mRNA. Ten micrograms of total RNA prepared from WT and KI mice hippocampi were analyzed in a formaldehyde-containing 1% agarose gel. The numbers on the left side indicate the sizes of the digoxigenin-labeled RNA molecular weight markers. Arrowheads indicate the major band for each mRNA.

    Techniques Used: Expressing, Western Blot, Quantitation Assay, Mutagenesis, Northern Blot, Agarose Gel Electrophoresis, Labeling, Molecular Weight

    rabbit anti psd 93  (Alomone Labs)


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    Alomone Labs rabbit anti psd 93
    Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and <t>PSD-93</t> knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.
    Rabbit Anti Psd 93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti psd 93 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein"

    Article Title: Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein

    Journal: Molecular Pain

    doi: 10.1186/1744-8069-4-45

    Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and PSD-93 knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.
    Figure Legend Snippet: Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and PSD-93 knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.

    Techniques Used: Expressing, Knock-Out, Western Blot

    Effect of targeted disruption of PSD-93 gene on morphine analgesic tolerance. (A) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following acute morphine (M) analgesic tolerance induced by two subcutaneous injections of morphine (100 mg/kg, 12 h apart). Control groups received saline (S) injection on the same schedule. (B) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following chronic morphine analgesic tolerance induced by subcutaneous injections of morphine (10 mg/kg) twice daily for 6 days. Control groups received saline injections on the same schedule. (C) Time course of morphine-induced analgesia and effect of MK-801 in WT and PSD-93 KO mice following subcutaneous injections of 10 mg/kg morphine twice daily plus intraperitoneal injection of saline or 0.3 mg/kg MK-801 once daily for 6 days. ** P < 0.01 vs the corresponding value on day 1.
    Figure Legend Snippet: Effect of targeted disruption of PSD-93 gene on morphine analgesic tolerance. (A) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following acute morphine (M) analgesic tolerance induced by two subcutaneous injections of morphine (100 mg/kg, 12 h apart). Control groups received saline (S) injection on the same schedule. (B) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following chronic morphine analgesic tolerance induced by subcutaneous injections of morphine (10 mg/kg) twice daily for 6 days. Control groups received saline injections on the same schedule. (C) Time course of morphine-induced analgesia and effect of MK-801 in WT and PSD-93 KO mice following subcutaneous injections of 10 mg/kg morphine twice daily plus intraperitoneal injection of saline or 0.3 mg/kg MK-801 once daily for 6 days. ** P < 0.01 vs the corresponding value on day 1.

    Techniques Used: Injection

    Effect of targeted disruption of PSD-93 gene on mechanical allodynia and thermal hyperalgesia after repeated morphine injection. WT and PSD-93 KO mice were injected twice daily with subcutaneous morphine (M, 20 mg/kg) and once daily with intraperitoneal saline (S) or 0.3 mg/kg MK-801 for 6 days. (A-D) Withdrawal responses of left (A and C) and right (B and D) hind paws to 0.24 mN (A and B) and 1.47 mN (C and D) intensity mechanical stimuli on days 1, 2, and 4 after morphine withdrawal. ** P < 0.01 vs the corresponding baseline. (E and F) Withdrawal response of left (E) and right (F) hind paws to thermal stimulation on days 1, 2, and 4 after morphine withdrawal. * P < 0.05 vs the corresponding baseline.
    Figure Legend Snippet: Effect of targeted disruption of PSD-93 gene on mechanical allodynia and thermal hyperalgesia after repeated morphine injection. WT and PSD-93 KO mice were injected twice daily with subcutaneous morphine (M, 20 mg/kg) and once daily with intraperitoneal saline (S) or 0.3 mg/kg MK-801 for 6 days. (A-D) Withdrawal responses of left (A and C) and right (B and D) hind paws to 0.24 mN (A and B) and 1.47 mN (C and D) intensity mechanical stimuli on days 1, 2, and 4 after morphine withdrawal. ** P < 0.01 vs the corresponding baseline. (E and F) Withdrawal response of left (E) and right (F) hind paws to thermal stimulation on days 1, 2, and 4 after morphine withdrawal. * P < 0.05 vs the corresponding baseline.

    Techniques Used: Injection

    Effect of targeted disruption of PSD-93 gene on physical dependence induced by repeated morphine injections. WT and KO mice were given morphine injections (10 mg/kg, s.c.) twice daily for 6 days plus a once-daily intraperitoneal injection of either saline or 0.3 mg/kg MK-801. Bars represent mean number of jumps in the 15-min period following a single naloxone injection (2 mg/kg) that was given 2 h after a final 10-mg/kg subcutaneous morphine injection. ** P < 0.01 vs the saline + morphine group.
    Figure Legend Snippet: Effect of targeted disruption of PSD-93 gene on physical dependence induced by repeated morphine injections. WT and KO mice were given morphine injections (10 mg/kg, s.c.) twice daily for 6 days plus a once-daily intraperitoneal injection of either saline or 0.3 mg/kg MK-801. Bars represent mean number of jumps in the 15-min period following a single naloxone injection (2 mg/kg) that was given 2 h after a final 10-mg/kg subcutaneous morphine injection. ** P < 0.01 vs the saline + morphine group.

    Techniques Used: Injection

    rabbit polyclonal anti psd 93  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti psd 93
    PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas <t>PSD-93</t> and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.
    Rabbit Polyclonal Anti Psd 93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti psd 93/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti psd 93 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "An altered glial phenotype in the NL3 R451C mouse model of autism"

    Article Title: An altered glial phenotype in the NL3 R451C mouse model of autism

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-71171-y

    PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas PSD-93 and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.
    Figure Legend Snippet: PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas PSD-93 and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.

    Techniques Used: Expressing, Western Blot, Two Tailed Test

    Antibodies used in Western blot analysis.
    Figure Legend Snippet: Antibodies used in Western blot analysis.

    Techniques Used: Western Blot

    anti psd 93 antibody  (Alomone Labs)


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    Alomone Labs anti psd 93 antibody
    Anti Psd 93 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti chapsyn110  (Alomone Labs)


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    psd 93  (Alomone Labs)


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    psd 93  (Alomone Labs)


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    Alomone Labs psd 93
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    anti psd93  (Alomone Labs)


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    Alomone Labs anti psd93
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    rabbit anti psd 93  (Alomone Labs)


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    Alomone Labs rabbit anti psd 93
    Efficacy of pCMV-U6-siRNA constructs. (A)RT-PCR analysis showed that a 495 bp product from <t>PSD-93</t> was inhibited significantly by siP3, GAPDH mRNA was used as a loading Control. siP3 significantly decreased the mRNA of PSD93 by 80.2% of control.(B) Confirmation by western blot analysis of silencing PSD93 expression, The blots were reprobed with anti-GAPDH antibody to verify protein loading. (C) Relative protein level of PSD-93 after siRNA transfection, PSD93 decreased by 79.5% compared to control group when transfected with siP3.
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    1) Product Images from "PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity"

    Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

    Journal:

    doi: 10.1016/j.neuroscience.2010.01.030

    Efficacy of pCMV-U6-siRNA constructs. (A)RT-PCR analysis showed that a 495 bp product from PSD-93 was inhibited significantly by siP3, GAPDH mRNA was used as a loading Control. siP3 significantly decreased the mRNA of PSD93 by 80.2% of control.(B) Confirmation by western blot analysis of silencing PSD93 expression, The blots were reprobed with anti-GAPDH antibody to verify protein loading. (C) Relative protein level of PSD-93 after siRNA transfection, PSD93 decreased by 79.5% compared to control group when transfected with siP3.
    Figure Legend Snippet: Efficacy of pCMV-U6-siRNA constructs. (A)RT-PCR analysis showed that a 495 bp product from PSD-93 was inhibited significantly by siP3, GAPDH mRNA was used as a loading Control. siP3 significantly decreased the mRNA of PSD93 by 80.2% of control.(B) Confirmation by western blot analysis of silencing PSD93 expression, The blots were reprobed with anti-GAPDH antibody to verify protein loading. (C) Relative protein level of PSD-93 after siRNA transfection, PSD93 decreased by 79.5% compared to control group when transfected with siP3.

    Techniques Used: Construct, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Transfection

    PSD-93 deficiency or knockdown protects against NMDA-stimulated neurotoxicity. Cortical neurons were cultured from WT and PSD-93 KO mice and treated with NMDA (0, 10, 20, 30, 40, and 60 μM) in the presence of 10 μM CNQX and 2 μM nimodipine. (A) Neuronal viability was assessed by MTT assay. (B) Percentage of neuronal death was determined by propidium iodide and calcein AM staining. * P < 0.05, ** P < 0.01 vs corresponding WT. n = 6 repeats. (C) Representative photographs showing propidium iodide and calcein AM staining of cortical neurons cultured from WT and PSD-93 KO mice treated with or without 30 μM NMDA or 10 μM MK-801+ 30 μM NMDA. Scale bar: 60 μm. (D) acute knock down PSD-93 by siP3 can inhibit the toxicity induced by NMDA at the dose of 30μM. MTT assay shows the cell viability reduced by 44.12% **P<0.01 vs normal group. However, siP3 can alleviate this toxicity, the cell viability increase 17%,# P<0.05 vs group transfected by siLuc.
    Figure Legend Snippet: PSD-93 deficiency or knockdown protects against NMDA-stimulated neurotoxicity. Cortical neurons were cultured from WT and PSD-93 KO mice and treated with NMDA (0, 10, 20, 30, 40, and 60 μM) in the presence of 10 μM CNQX and 2 μM nimodipine. (A) Neuronal viability was assessed by MTT assay. (B) Percentage of neuronal death was determined by propidium iodide and calcein AM staining. * P < 0.05, ** P < 0.01 vs corresponding WT. n = 6 repeats. (C) Representative photographs showing propidium iodide and calcein AM staining of cortical neurons cultured from WT and PSD-93 KO mice treated with or without 30 μM NMDA or 10 μM MK-801+ 30 μM NMDA. Scale bar: 60 μm. (D) acute knock down PSD-93 by siP3 can inhibit the toxicity induced by NMDA at the dose of 30μM. MTT assay shows the cell viability reduced by 44.12% **P<0.01 vs normal group. However, siP3 can alleviate this toxicity, the cell viability increase 17%,# P<0.05 vs group transfected by siLuc.

    Techniques Used: Cell Culture, MTT Assay, Staining, Transfection

    PSD-93 deficiency has no effect on non-NMDA receptor-triggered neurotoxicity in cultured cortical neurons. Cultured neurons were treated with kainate at the doses shown in the presence of 10 μM MK-801 and 2 μM nimodipine. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). n = 6 repeats.
    Figure Legend Snippet: PSD-93 deficiency has no effect on non-NMDA receptor-triggered neurotoxicity in cultured cortical neurons. Cultured neurons were treated with kainate at the doses shown in the presence of 10 μM MK-801 and 2 μM nimodipine. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). n = 6 repeats.

    Techniques Used: Cell Culture, Staining, MTT Assay

    MK-801 attenuates NMDA-induced neurotoxicity in cultured cortical neurons from WT but not from PSD-93 KO mice. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). * P < 0.05 vs the corresponding NMDA concentration alone. n = 6 repeats.
    Figure Legend Snippet: MK-801 attenuates NMDA-induced neurotoxicity in cultured cortical neurons from WT but not from PSD-93 KO mice. Percentage of neuronal death was determined by propidium iodide and calcein AM staining (top) and neuronal viability by MTT assay (bottom). * P < 0.05 vs the corresponding NMDA concentration alone. n = 6 repeats.

    Techniques Used: Cell Culture, Staining, MTT Assay, Concentration Assay

    PSD-93 deficiency significantly reduces synaptic expression of NR2A and NR2B in cultured cortical neurons. (A) Representative Western blots showing the levels of PSD-93, PSD-95, NR2A, and NR2B in the total soluble and synaptosomal fractions. (B) Statistical summary of the densitometric analysis expressed relative to WT mice after normalization to corresponding β-actin or N-cadherin. ** P < 0.01 vs WT mice. n = 3 repeats.
    Figure Legend Snippet: PSD-93 deficiency significantly reduces synaptic expression of NR2A and NR2B in cultured cortical neurons. (A) Representative Western blots showing the levels of PSD-93, PSD-95, NR2A, and NR2B in the total soluble and synaptosomal fractions. (B) Statistical summary of the densitometric analysis expressed relative to WT mice after normalization to corresponding β-actin or N-cadherin. ** P < 0.01 vs WT mice. n = 3 repeats.

    Techniques Used: Expressing, Cell Culture, Western Blot

    PSD-93 deficiency attenuates NMDA-stimulated Ca2+ loading in cultured cortical neurons. Cultures were challenged for 5 or 10 min with 30 μM NMDA, 10 μM CNQX, and 2 μM nimodipine in medium containing 45CaCl2. CMP: counts per minute. * P < 0.05 vs corresponding control. n = 6 repeats.
    Figure Legend Snippet: PSD-93 deficiency attenuates NMDA-stimulated Ca2+ loading in cultured cortical neurons. Cultures were challenged for 5 or 10 min with 30 μM NMDA, 10 μM CNQX, and 2 μM nimodipine in medium containing 45CaCl2. CMP: counts per minute. * P < 0.05 vs corresponding control. n = 6 repeats.

    Techniques Used: Cell Culture

    PSD-93 deficiency decreases NMDA-stimulated increase in cGMP in cultured cortical neurons. (A) NMDA-induced neurotoxicity was NOS-dependent in cultured cortical neurons. Cultured neurons were treated with NMDA (30 μM or 60 μM) with or without L-NAME (10 μM or 30 μM). Neuronal viability was determined by MTT assay (top) and percentage of neuronal death by propidium iodide and calcein AM staining (bottom). * P < 0.05 vs NMDA alone. n = 6 repeats. (B) Cultured neurons from WT and PSD-93 KO mice were exposed to 30 μM NMDA. PSD-93 deficiency decreased the NMDA-stimulated increase in cGMP level in cultured cortical neurons. * P < 0.05 vs the corresponding NMDA-treated group from WT mice. n = 6 repeats.
    Figure Legend Snippet: PSD-93 deficiency decreases NMDA-stimulated increase in cGMP in cultured cortical neurons. (A) NMDA-induced neurotoxicity was NOS-dependent in cultured cortical neurons. Cultured neurons were treated with NMDA (30 μM or 60 μM) with or without L-NAME (10 μM or 30 μM). Neuronal viability was determined by MTT assay (top) and percentage of neuronal death by propidium iodide and calcein AM staining (bottom). * P < 0.05 vs NMDA alone. n = 6 repeats. (B) Cultured neurons from WT and PSD-93 KO mice were exposed to 30 μM NMDA. PSD-93 deficiency decreased the NMDA-stimulated increase in cGMP level in cultured cortical neurons. * P < 0.05 vs the corresponding NMDA-treated group from WT mice. n = 6 repeats.

    Techniques Used: Cell Culture, MTT Assay, Staining

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    • rabbit anti psd93  (Alomone Labs)
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    Alomone Labs rabbit anti psd93
    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for <t>PSD93.</t> B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.
    Rabbit Anti Psd93, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti psd 93  (Alomone Labs)
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    Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: <t>PSD-93,</t> 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.
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    Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and <t>PSD-93</t> knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.
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    PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas <t>PSD-93</t> and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.
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    A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.

    Journal: PLoS ONE

    Article Title: Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation

    doi: 10.1371/journal.pone.0023978

    Figure Lengend Snippet: A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.

    Article Snippet: Rabbit phosphor-specific anti-Src (pY418), recognizing both Y423 in mouse Src and Y419 in mouse Fyn, as well as anti-Src (pY529), recognizing both Y534 in mouse Src and Y530 in mouse Fyn due to the highly conserved sequence of these residues were purchased from BioSource, rabbit anti-Csk (C-20) and anti-Ctk from Santa Cruz, mouse-anti-Csk (clone 52) and mouse anti-CHK from BD BioSciences, mouse anti-Src (clone GD11) from Upstate (Lake Placid, NY), rabbit anti-PSD93 from Alomone Labs Ltd., rabbit anti-c-Src (Y215) from ECM Biosciences, and mouse anti-β-actin (clone AC-15) from Sigma.

    Techniques: Western Blot, Staining, Lysis, Centrifugation, Binding Assay, Dot Blot

    Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: PSD-93, 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.

    Journal: Molecular Brain

    Article Title: Impaired synaptic clustering of postsynaptic density proteins and altered signal transmission in hippocampal neurons, and disrupted learning behavior in PDZ1 and PDZ2 ligand binding-deficient PSD-95 knockin mice

    doi: 10.1186/1756-6606-5-43

    Figure Lengend Snippet: Altered PSD protein composition in the hippocampal PSD fraction of KI mice during development. A , Immunoblot analyses of PSD fractions from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of WT and KI PSD proteins were analyzed per lane: PSD-93, 0.5 μg; SAP102, 0.5 μg; SAP97, 2 μg; GluN2A, 0.5 μg; GluN2B, 0.5 μg; GluA1, 0.5 μg; GluA2/3, 0.5 μg; and γ2/4/8, 2 μg. The arrowhead indicates the degraded bands of PSD-93. For PSD-95, 0.25 μg of WT and 0.5 μg of KI PSD proteins were analyzed for comparison on identical membrane filters and their band intensities were corrected in B . B , Quantitation of various proteins in the PSD fractions based on the results shown in A , normalized to the wild-type level at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of PSD fractions at various ages (P14–P65) were examined. Simple comparison of the means and SEM of WT and KI data was performed using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD fraction, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, GluA1, GluA2/3, and γ2/4/8, and increased levels of SAP102 during development are observed compared to the WT mice.

    Article Snippet: The following antibodies were used: anti-PSD-95 (mouse monoclonal; K28/43; 1:1000; NeuroMab), anti-PSD-93 (rabbit polyclonal; 1:200; Alomone Labs), anti-SAP97 (mouse monoclonal; K64/15; 1:200; NeuroMab), anti-GluA1 (rabbit polyclonal; 1:400; Chemicon), anti-GluA2/3 (rabbit polyclonal; 1:500; Chemicon), anti-GluN2A (mouse monoclonal; 1:500; BD Transduction Laboratories), anti-GluN2B (mouse monoclonal; 1:250; BD Transduction Laboratories), anti-TARPGamma2/4/8 (mouse monoclonal; N245/36; 1:200; NeuroMab), and anti-SynGAP (rabbit polyclonal; 1:1000; Affinity BioReagents), anti-AKAP150 (rabbit polyclonal; 1:1000; Millipore), anti-pan-Shank (mouse monoclonal; 1:1000; StressMarq Biosciences), anti-mGluR5 (rabbit polyclonal; 1:2000; Millipore), anti-Homer antiserum (rabbit polyclonal; 1:10000).

    Techniques: Western Blot, Quantitation Assay, Mutagenesis

    Altered expression of SAP102 and PSD-93 in the hippocampus of KI mice. A , Immunoblot analyses of hippocampal homogenates from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of homogenate proteins were analyzed per lane: PSD-93 and SAP102, WT and KI: 5 μg; GluA1 and GluA2/3, WT and KI: 10 μg; and PSD-95, WT: 5 μg, KI: 20 μg. The arrowhead indicates the degraded bands of PSD-93. B , Quantitation of PSD proteins in the hippocampal homogenates based on the results shown in A , normalized to the wild-type levels at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of hippocampal homogenates at various ages (P4–P65) were examined. The means and SEM of WT and KI data were compared using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD homogenates, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, and increased levels of SAP102 were observed during development compared to the WT mice, while the levels of GluA1 and GluA2/3 were not significantly different from those of the WT mice. C , Northern blot analyses of PSD-95 and SAP102 mRNA. Ten micrograms of total RNA prepared from WT and KI mice hippocampi were analyzed in a formaldehyde-containing 1% agarose gel. The numbers on the left side indicate the sizes of the digoxigenin-labeled RNA molecular weight markers. Arrowheads indicate the major band for each mRNA.

    Journal: Molecular Brain

    Article Title: Impaired synaptic clustering of postsynaptic density proteins and altered signal transmission in hippocampal neurons, and disrupted learning behavior in PDZ1 and PDZ2 ligand binding-deficient PSD-95 knockin mice

    doi: 10.1186/1756-6606-5-43

    Figure Lengend Snippet: Altered expression of SAP102 and PSD-93 in the hippocampus of KI mice. A , Immunoblot analyses of hippocampal homogenates from WT and KI mice at the respective developmental ages for the indicated proteins. The following amounts of homogenate proteins were analyzed per lane: PSD-93 and SAP102, WT and KI: 5 μg; GluA1 and GluA2/3, WT and KI: 10 μg; and PSD-95, WT: 5 μg, KI: 20 μg. The arrowhead indicates the degraded bands of PSD-93. B , Quantitation of PSD proteins in the hippocampal homogenates based on the results shown in A , normalized to the wild-type levels at P30 (set as 100%). The histograms show the mean ± SEM (open bars, WT; filled bars, KI). Three sets of hippocampal homogenates at various ages (P4–P65) were examined. The means and SEM of WT and KI data were compared using Student’s t-test. * P < 0.05, ** P < 0.005, *** P < 0.001. In the KI PSD homogenates, extensively low levels of mutant PSD-95, significantly decreased levels of PSD-93, and increased levels of SAP102 were observed during development compared to the WT mice, while the levels of GluA1 and GluA2/3 were not significantly different from those of the WT mice. C , Northern blot analyses of PSD-95 and SAP102 mRNA. Ten micrograms of total RNA prepared from WT and KI mice hippocampi were analyzed in a formaldehyde-containing 1% agarose gel. The numbers on the left side indicate the sizes of the digoxigenin-labeled RNA molecular weight markers. Arrowheads indicate the major band for each mRNA.

    Article Snippet: The following antibodies were used: anti-PSD-95 (mouse monoclonal; K28/43; 1:1000; NeuroMab), anti-PSD-93 (rabbit polyclonal; 1:200; Alomone Labs), anti-SAP97 (mouse monoclonal; K64/15; 1:200; NeuroMab), anti-GluA1 (rabbit polyclonal; 1:400; Chemicon), anti-GluA2/3 (rabbit polyclonal; 1:500; Chemicon), anti-GluN2A (mouse monoclonal; 1:500; BD Transduction Laboratories), anti-GluN2B (mouse monoclonal; 1:250; BD Transduction Laboratories), anti-TARPGamma2/4/8 (mouse monoclonal; N245/36; 1:200; NeuroMab), and anti-SynGAP (rabbit polyclonal; 1:1000; Affinity BioReagents), anti-AKAP150 (rabbit polyclonal; 1:1000; Millipore), anti-pan-Shank (mouse monoclonal; 1:1000; StressMarq Biosciences), anti-mGluR5 (rabbit polyclonal; 1:2000; Millipore), anti-Homer antiserum (rabbit polyclonal; 1:10000).

    Techniques: Expressing, Western Blot, Quantitation Assay, Mutagenesis, Northern Blot, Agarose Gel Electrophoresis, Labeling, Molecular Weight

    Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and PSD-93 knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.

    Journal: Molecular Pain

    Article Title: Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein

    doi: 10.1186/1744-8069-4-45

    Figure Lengend Snippet: Expression of NR2A and NR2B in total soluble (A) and synaptosomal membrane (B) fractions from dorsal horn, forebrain cortex, and cerebellum in wild type (WT) and PSD-93 knockout (KO) mice. (A) Top: representative Western blots showing the levels of PSD-93, NR2A, and NR2B in the total soluble fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control (β-actin). (B) Top: representative Western blots showing the amounts of PSD-93, NR2A, and NR2B in the synaptosomal membrane fraction. Bottom: statistical summary of the densitometric analysis expressed relative to the corresponding loading control ( N -cadherin). ** P < 0.01 vs the corresponding naïve WT mice.

    Article Snippet: The blotting membrane was blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti- N -cadherin (1: 1,000; BD Biosciences, Palo Alto, CA), rabbit anti-NR2A (1: 200, Upstate/CHEMICON, Temecula, CA), rabbit anti-NR2B (1:500, Upstate/CHEMICON), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

    Techniques: Expressing, Knock-Out, Western Blot

    Effect of targeted disruption of PSD-93 gene on morphine analgesic tolerance. (A) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following acute morphine (M) analgesic tolerance induced by two subcutaneous injections of morphine (100 mg/kg, 12 h apart). Control groups received saline (S) injection on the same schedule. (B) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following chronic morphine analgesic tolerance induced by subcutaneous injections of morphine (10 mg/kg) twice daily for 6 days. Control groups received saline injections on the same schedule. (C) Time course of morphine-induced analgesia and effect of MK-801 in WT and PSD-93 KO mice following subcutaneous injections of 10 mg/kg morphine twice daily plus intraperitoneal injection of saline or 0.3 mg/kg MK-801 once daily for 6 days. ** P < 0.01 vs the corresponding value on day 1.

    Journal: Molecular Pain

    Article Title: Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein

    doi: 10.1186/1744-8069-4-45

    Figure Lengend Snippet: Effect of targeted disruption of PSD-93 gene on morphine analgesic tolerance. (A) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following acute morphine (M) analgesic tolerance induced by two subcutaneous injections of morphine (100 mg/kg, 12 h apart). Control groups received saline (S) injection on the same schedule. (B) The cumulative dose-response curves of morphine in WT and PSD-93 KO mice following chronic morphine analgesic tolerance induced by subcutaneous injections of morphine (10 mg/kg) twice daily for 6 days. Control groups received saline injections on the same schedule. (C) Time course of morphine-induced analgesia and effect of MK-801 in WT and PSD-93 KO mice following subcutaneous injections of 10 mg/kg morphine twice daily plus intraperitoneal injection of saline or 0.3 mg/kg MK-801 once daily for 6 days. ** P < 0.01 vs the corresponding value on day 1.

    Article Snippet: The blotting membrane was blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti- N -cadherin (1: 1,000; BD Biosciences, Palo Alto, CA), rabbit anti-NR2A (1: 200, Upstate/CHEMICON, Temecula, CA), rabbit anti-NR2B (1:500, Upstate/CHEMICON), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

    Techniques: Injection

    Effect of targeted disruption of PSD-93 gene on mechanical allodynia and thermal hyperalgesia after repeated morphine injection. WT and PSD-93 KO mice were injected twice daily with subcutaneous morphine (M, 20 mg/kg) and once daily with intraperitoneal saline (S) or 0.3 mg/kg MK-801 for 6 days. (A-D) Withdrawal responses of left (A and C) and right (B and D) hind paws to 0.24 mN (A and B) and 1.47 mN (C and D) intensity mechanical stimuli on days 1, 2, and 4 after morphine withdrawal. ** P < 0.01 vs the corresponding baseline. (E and F) Withdrawal response of left (E) and right (F) hind paws to thermal stimulation on days 1, 2, and 4 after morphine withdrawal. * P < 0.05 vs the corresponding baseline.

    Journal: Molecular Pain

    Article Title: Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein

    doi: 10.1186/1744-8069-4-45

    Figure Lengend Snippet: Effect of targeted disruption of PSD-93 gene on mechanical allodynia and thermal hyperalgesia after repeated morphine injection. WT and PSD-93 KO mice were injected twice daily with subcutaneous morphine (M, 20 mg/kg) and once daily with intraperitoneal saline (S) or 0.3 mg/kg MK-801 for 6 days. (A-D) Withdrawal responses of left (A and C) and right (B and D) hind paws to 0.24 mN (A and B) and 1.47 mN (C and D) intensity mechanical stimuli on days 1, 2, and 4 after morphine withdrawal. ** P < 0.01 vs the corresponding baseline. (E and F) Withdrawal response of left (E) and right (F) hind paws to thermal stimulation on days 1, 2, and 4 after morphine withdrawal. * P < 0.05 vs the corresponding baseline.

    Article Snippet: The blotting membrane was blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti- N -cadherin (1: 1,000; BD Biosciences, Palo Alto, CA), rabbit anti-NR2A (1: 200, Upstate/CHEMICON, Temecula, CA), rabbit anti-NR2B (1:500, Upstate/CHEMICON), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

    Techniques: Injection

    Effect of targeted disruption of PSD-93 gene on physical dependence induced by repeated morphine injections. WT and KO mice were given morphine injections (10 mg/kg, s.c.) twice daily for 6 days plus a once-daily intraperitoneal injection of either saline or 0.3 mg/kg MK-801. Bars represent mean number of jumps in the 15-min period following a single naloxone injection (2 mg/kg) that was given 2 h after a final 10-mg/kg subcutaneous morphine injection. ** P < 0.01 vs the saline + morphine group.

    Journal: Molecular Pain

    Article Title: Distinct expression of synaptic NR2A and NR2B in the central nervous system and impaired morphine tolerance and physical dependence in mice deficient in postsynaptic density-93 protein

    doi: 10.1186/1744-8069-4-45

    Figure Lengend Snippet: Effect of targeted disruption of PSD-93 gene on physical dependence induced by repeated morphine injections. WT and KO mice were given morphine injections (10 mg/kg, s.c.) twice daily for 6 days plus a once-daily intraperitoneal injection of either saline or 0.3 mg/kg MK-801. Bars represent mean number of jumps in the 15-min period following a single naloxone injection (2 mg/kg) that was given 2 h after a final 10-mg/kg subcutaneous morphine injection. ** P < 0.01 vs the saline + morphine group.

    Article Snippet: The blotting membrane was blocked with 3% non-fat dry milk for 1 h and incubated overnight at 4°C with rabbit anti-PSD-93 (1:1,000; Alomone Labs Ltd, Jerusalem, Israel), rabbit anti- N -cadherin (1: 1,000; BD Biosciences, Palo Alto, CA), rabbit anti-NR2A (1: 200, Upstate/CHEMICON, Temecula, CA), rabbit anti-NR2B (1:500, Upstate/CHEMICON), or monoclonal mouse anti-β-actin (1:10,000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

    Techniques: Injection

    PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas PSD-93 and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.

    Journal: Scientific Reports

    Article Title: An altered glial phenotype in the NL3 R451C mouse model of autism

    doi: 10.1038/s41598-020-71171-y

    Figure Lengend Snippet: PSD-95 protein levels are increased in the striatum of NL3 R451C mice, whereas PSD-93 and NL2 expression is unchanged. Levels of postsynaptic proteins, PSD-95, PSD-93 and NL2 in cortical ( A , G , M ), striatal ( C , I , O ) and cerebellar ( E , K , Q ) brain lysates from NL3 R451C and WT mice were analysed via Western blot. Densitometric analysis was performed to demonstrate quantitative expression of PSD-95 ( B , D , F ), PSD-93 ( H , J , L ) and NL2 ( N , P , R ) relative to β actin expression. Genotype differences were analysed using an unpaired, two-tailed Student’s t-test (n = 6 mice in each group); *p < 0.05. Data represented as mean ± SEM.

    Article Snippet: Rabbit polyclonal anti-PSD-93 (110 kDa) , Alomone Labs #APZ002 , 1:500.

    Techniques: Expressing, Western Blot, Two Tailed Test

    Antibodies used in Western blot analysis.

    Journal: Scientific Reports

    Article Title: An altered glial phenotype in the NL3 R451C mouse model of autism

    doi: 10.1038/s41598-020-71171-y

    Figure Lengend Snippet: Antibodies used in Western blot analysis.

    Article Snippet: Rabbit polyclonal anti-PSD-93 (110 kDa) , Alomone Labs #APZ002 , 1:500.

    Techniques: Western Blot