glun2a  (Alomone Labs)


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    Structured Review

    Alomone Labs glun2a
    Synaptic GluN2B-NMDAR are laterally redistributed in the synaptic area following chem LTP in immature neurons Detection of a single QD (30-Hz acquisition) in our experimental conditions. The high signal-to-noise ratio (SNR) ( > 5) enables the detection and location of the signal with a high pointing accuracy (˜30 nm). The QD fluorescence is quantified on a pseudocolor scale (low: red; high: yellow). Scale bar = 800 nm. A 500-frame stack is obtained while tracking down a single NMDAR/QD complex. On each frame, a single GluN2B- (green) or <t>GluN2A-</t> (blue) QD particle complex is detected and precisely located within synaptic (dark gray) and perisynaptic (320-nm annulus around the synapse; light gray) areas. Those 500 locations are then projected on a single image, providing the successive positions of this receptor/particle complex during the 500-frame stack. Note that GluN2A-NMDAR are more concentrated within the core of the PSD. Scale bar image = 300 nm; synaptic areas = 200 nm. Relative fraction of synaptic and perisynaptic GluN2-QD particles, calculated before and after chem LTP for both GluN2B- (left) and GluN2A-NMDAR (right) ( n = 25 and 20 dendritic fields before and after chem LTP, respectively). Note the significant decrease in the relative synaptic content in GluN2B-NMDAR particles right after chem LTP (Student's t -test, ** P
    Glun2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glun2a/product/Alomone Labs
    Average 95 stars, based on 7 article reviews
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    Images

    1) Product Images from "Surface dynamics of GluN2B-NMDA receptors controls plasticity of maturing glutamate synapses"

    Article Title: Surface dynamics of GluN2B-NMDA receptors controls plasticity of maturing glutamate synapses

    Journal: The EMBO Journal

    doi: 10.1002/embj.201386356

    Synaptic GluN2B-NMDAR are laterally redistributed in the synaptic area following chem LTP in immature neurons Detection of a single QD (30-Hz acquisition) in our experimental conditions. The high signal-to-noise ratio (SNR) ( > 5) enables the detection and location of the signal with a high pointing accuracy (˜30 nm). The QD fluorescence is quantified on a pseudocolor scale (low: red; high: yellow). Scale bar = 800 nm. A 500-frame stack is obtained while tracking down a single NMDAR/QD complex. On each frame, a single GluN2B- (green) or GluN2A- (blue) QD particle complex is detected and precisely located within synaptic (dark gray) and perisynaptic (320-nm annulus around the synapse; light gray) areas. Those 500 locations are then projected on a single image, providing the successive positions of this receptor/particle complex during the 500-frame stack. Note that GluN2A-NMDAR are more concentrated within the core of the PSD. Scale bar image = 300 nm; synaptic areas = 200 nm. Relative fraction of synaptic and perisynaptic GluN2-QD particles, calculated before and after chem LTP for both GluN2B- (left) and GluN2A-NMDAR (right) ( n = 25 and 20 dendritic fields before and after chem LTP, respectively). Note the significant decrease in the relative synaptic content in GluN2B-NMDAR particles right after chem LTP (Student's t -test, ** P
    Figure Legend Snippet: Synaptic GluN2B-NMDAR are laterally redistributed in the synaptic area following chem LTP in immature neurons Detection of a single QD (30-Hz acquisition) in our experimental conditions. The high signal-to-noise ratio (SNR) ( > 5) enables the detection and location of the signal with a high pointing accuracy (˜30 nm). The QD fluorescence is quantified on a pseudocolor scale (low: red; high: yellow). Scale bar = 800 nm. A 500-frame stack is obtained while tracking down a single NMDAR/QD complex. On each frame, a single GluN2B- (green) or GluN2A- (blue) QD particle complex is detected and precisely located within synaptic (dark gray) and perisynaptic (320-nm annulus around the synapse; light gray) areas. Those 500 locations are then projected on a single image, providing the successive positions of this receptor/particle complex during the 500-frame stack. Note that GluN2A-NMDAR are more concentrated within the core of the PSD. Scale bar image = 300 nm; synaptic areas = 200 nm. Relative fraction of synaptic and perisynaptic GluN2-QD particles, calculated before and after chem LTP for both GluN2B- (left) and GluN2A-NMDAR (right) ( n = 25 and 20 dendritic fields before and after chem LTP, respectively). Note the significant decrease in the relative synaptic content in GluN2B-NMDAR particles right after chem LTP (Student's t -test, ** P

    Techniques Used: Fluorescence

    2) Product Images from "Human endogenous retroviral protein triggers deficit in glutamate synapse maturation and behaviors associated with psychosis"

    Article Title: Human endogenous retroviral protein triggers deficit in glutamate synapse maturation and behaviors associated with psychosis

    Journal: Science Advances

    doi: 10.1126/sciadv.abc0708

    Recombinant Env disperses synaptic NMDAR in hippocampal neurons without interfering with the ionotropic function. ( A ) Experimental setup of (yellow) neurons [microtubule-associated protein 2 (MAP-2) positive] and (magenta) glia [glial fibrillary acidic protein (GFAP) and Iba1 positive]. Representative NMDAR-mediated Ca 2+ signals and detected transients (dots) 5 min after Cont. (vehicle) or Env exposure. Scale bars, 30 and 1 μm. ( B ) Ca 2+ frequency ratio (post/pre) for blank ( n = 47/5 spines/neurons); Cont. ( n = 92/7); and Env: 0.5 μg/ml ( n = 87/6), 1.0 μg/ml ( n = 114/9), and 10 μg/ml ( n = 18/1). ( C ) Representative NMDA traces and mean peak amplitudes before (pre) and 5 min after (post) Cont. or Env application. ( D ) Representative trajectories of GluN2A- and GluN2B-NMDAR-QD complexes at postsynaptic densities (PSD). Scale bar, 1 μm. ( E ) Synaptic increase of GluN2B-NMDAR but not GluN2A-NMDAR surface diffusion. Data are normalized to pre-exposure for individual neurons. GluN2A, Cont. (vehicle; n = 176/4 trajectories/neuron) and Env ( n = 200/5). GluN2B, Cont. ( n = 670/16) and Env ( n = 792/17). *** P
    Figure Legend Snippet: Recombinant Env disperses synaptic NMDAR in hippocampal neurons without interfering with the ionotropic function. ( A ) Experimental setup of (yellow) neurons [microtubule-associated protein 2 (MAP-2) positive] and (magenta) glia [glial fibrillary acidic protein (GFAP) and Iba1 positive]. Representative NMDAR-mediated Ca 2+ signals and detected transients (dots) 5 min after Cont. (vehicle) or Env exposure. Scale bars, 30 and 1 μm. ( B ) Ca 2+ frequency ratio (post/pre) for blank ( n = 47/5 spines/neurons); Cont. ( n = 92/7); and Env: 0.5 μg/ml ( n = 87/6), 1.0 μg/ml ( n = 114/9), and 10 μg/ml ( n = 18/1). ( C ) Representative NMDA traces and mean peak amplitudes before (pre) and 5 min after (post) Cont. or Env application. ( D ) Representative trajectories of GluN2A- and GluN2B-NMDAR-QD complexes at postsynaptic densities (PSD). Scale bar, 1 μm. ( E ) Synaptic increase of GluN2B-NMDAR but not GluN2A-NMDAR surface diffusion. Data are normalized to pre-exposure for individual neurons. GluN2A, Cont. (vehicle; n = 176/4 trajectories/neuron) and Env ( n = 200/5). GluN2B, Cont. ( n = 670/16) and Env ( n = 792/17). *** P

    Techniques Used: Recombinant, Diffusion-based Assay

    Neonatal Env expression tunes glutamatergic synapse maturation and is necessary for psychotic-like behavior. ( A ) Subcellular fractionation of hippocampal tissue. Note the enrichment of NMDA receptors (represented by GluN2A) and postsynaptic density proteins (PSD-95) in P2 and synaptosome (synaptic proteins enriched) fractions compared to initial homogenate and the depletion of glia (GFAP) in the same. ( B ) Representative images of inserted gene distribution at P7. Scale bars, 1 mm. ( C ) Hippocampal inserted gene expression at ~P65. ( D ) Env exposure tends to influence GluN2A/PSD-95 subunit stabilization in the synapse (P7; Cont. versus Env; P = 0.06) that then remains constant. Interaction: F (1,29) = 8.69, two-way ANOVA and Bonferroni's multiple comparisons test; * P = 0.021 and *** P = 0.0004. ( E ) GluN2B/PSD-95 expression is unaffected by Env at both ages. Interaction: F (1,29) = 2.46, P = 0.13, two-way ANOVA and Bonferroni's multiple comparisons test; *** P
    Figure Legend Snippet: Neonatal Env expression tunes glutamatergic synapse maturation and is necessary for psychotic-like behavior. ( A ) Subcellular fractionation of hippocampal tissue. Note the enrichment of NMDA receptors (represented by GluN2A) and postsynaptic density proteins (PSD-95) in P2 and synaptosome (synaptic proteins enriched) fractions compared to initial homogenate and the depletion of glia (GFAP) in the same. ( B ) Representative images of inserted gene distribution at P7. Scale bars, 1 mm. ( C ) Hippocampal inserted gene expression at ~P65. ( D ) Env exposure tends to influence GluN2A/PSD-95 subunit stabilization in the synapse (P7; Cont. versus Env; P = 0.06) that then remains constant. Interaction: F (1,29) = 8.69, two-way ANOVA and Bonferroni's multiple comparisons test; * P = 0.021 and *** P = 0.0004. ( E ) GluN2B/PSD-95 expression is unaffected by Env at both ages. Interaction: F (1,29) = 2.46, P = 0.13, two-way ANOVA and Bonferroni's multiple comparisons test; *** P

    Techniques Used: Expressing, Fractionation

    3) Product Images from "Cholesterol modulates presynaptic and postsynaptic properties of excitatory synaptic transmission"

    Article Title: Cholesterol modulates presynaptic and postsynaptic properties of excitatory synaptic transmission

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-69454-5

    Cholesterol depletion reduces synaptic localization of NMDARs. ( A , C ) Colocalization of surface GluN2A (A, green) or GluN2B (C, green) and the postsynaptic marker Shank (red) in control and cholesterol-depleted neurons (10 mM MβCD pretreatment, 5 min). Scale bar 2 µm. ( B , D ) Bar graphs showing Pearson's coefficient for the colocalization indicate the reduction of synaptic localization of GluN2A and GluN2B after cholesterol depletion. ( E ) Colocalization of surface GluA1 (green) and the postsynaptic marker Shank (red) in control and cholesterol-depleted neurons (MβCD). Scale bar 2 µm. ( F ) Bar graph showing Pearson's coefficient for the colocalization. ( G ) Examples of typical dual AMPAR-NMDAR mEPSCs in control autaptic neurons having various AMPAR to NMDAR ratio. ( H ) Examples of typical dual AMPAR-NMDAR mEPSCs in 10 mM MβCD-pretreated autaptic neurons. ( I ) Examples of NMDAR mEPSCs obtained from average dual mEPSCs after AMPAR mEPSC subtraction. A control neuron (top trace) and a cholesterol-depleted neuron (bottom trace). The arrows indicate mEPSC onsets. ( J ) The comparison of average amplitude of NMDAR mEPSCs in control neurons and in cholesterol-depleted neurons. (* p
    Figure Legend Snippet: Cholesterol depletion reduces synaptic localization of NMDARs. ( A , C ) Colocalization of surface GluN2A (A, green) or GluN2B (C, green) and the postsynaptic marker Shank (red) in control and cholesterol-depleted neurons (10 mM MβCD pretreatment, 5 min). Scale bar 2 µm. ( B , D ) Bar graphs showing Pearson's coefficient for the colocalization indicate the reduction of synaptic localization of GluN2A and GluN2B after cholesterol depletion. ( E ) Colocalization of surface GluA1 (green) and the postsynaptic marker Shank (red) in control and cholesterol-depleted neurons (MβCD). Scale bar 2 µm. ( F ) Bar graph showing Pearson's coefficient for the colocalization. ( G ) Examples of typical dual AMPAR-NMDAR mEPSCs in control autaptic neurons having various AMPAR to NMDAR ratio. ( H ) Examples of typical dual AMPAR-NMDAR mEPSCs in 10 mM MβCD-pretreated autaptic neurons. ( I ) Examples of NMDAR mEPSCs obtained from average dual mEPSCs after AMPAR mEPSC subtraction. A control neuron (top trace) and a cholesterol-depleted neuron (bottom trace). The arrows indicate mEPSC onsets. ( J ) The comparison of average amplitude of NMDAR mEPSCs in control neurons and in cholesterol-depleted neurons. (* p

    Techniques Used: Marker

    Cholesterol depletion reduces the fraction of synaptic immobile NMDARs. ( A ) Surface NMDARs were detected using a QD-antibody complex directed against extracellular epitopes in GluN2A or GluN2B. Left, representative summed trajectories of NMDAR-QDs (red) acquired over a period of 25 s (20 Hz frame rate) in hippocampal neurons. Scale bar 5 µm. Right, representative examples of NMDAR reconstructed trajectories. ( B , C ) Diffusion coefficients for synaptic GluN2A-containing NMDARs and GluN2B-containing NMDARs in control and after cholesterol depletion (10 mM MβCD pretreatment, 5 min). ( D , E ) Diffusion coefficients for extrasynaptic GluN2A-containing NMDAR and GluN2B-containing NMDARs in control and after cholesterol depletion. ( F , G ) Diffusion coefficients for the mobile fraction of synaptic GluN2A and GluN2B-containing NMDARs in control and after cholesterol depletion. ( H , I ) Fraction of synaptic immobile receptors in control and after cholesterol depletion. (* p
    Figure Legend Snippet: Cholesterol depletion reduces the fraction of synaptic immobile NMDARs. ( A ) Surface NMDARs were detected using a QD-antibody complex directed against extracellular epitopes in GluN2A or GluN2B. Left, representative summed trajectories of NMDAR-QDs (red) acquired over a period of 25 s (20 Hz frame rate) in hippocampal neurons. Scale bar 5 µm. Right, representative examples of NMDAR reconstructed trajectories. ( B , C ) Diffusion coefficients for synaptic GluN2A-containing NMDARs and GluN2B-containing NMDARs in control and after cholesterol depletion (10 mM MβCD pretreatment, 5 min). ( D , E ) Diffusion coefficients for extrasynaptic GluN2A-containing NMDAR and GluN2B-containing NMDARs in control and after cholesterol depletion. ( F , G ) Diffusion coefficients for the mobile fraction of synaptic GluN2A and GluN2B-containing NMDARs in control and after cholesterol depletion. ( H , I ) Fraction of synaptic immobile receptors in control and after cholesterol depletion. (* p

    Techniques Used: Diffusion-based Assay

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    Alomone Labs nr2a
    CRIPT knockdown leads to a selective reduction in the abundance of GluA1 and SAP97. Mixed spinal cord cultures were infected with HSV engineered to express a miRNA targeting CRIPT or a scrambled control. Two days later, lysates were prepared and subjected to Western blottings. No more than six independent experiments were performed for the quantitative image analysis. CRIPT knockdown leads to a reduction in GluA1 and SAP97 abundance and no effect on the abundance of GluA2, GluA4, NR1, <t>NR2A,</t> NR2B, or PSD95. Representative images of Western blottings with actin loading controls are shown and quantification of band intensity in the bar graphs below; *significant difference between groups, p
    Nr2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Alomone Labs anti rabbit nr2a
    Assessment of <t>NR2A</t> and NR2B expression in the PrV by Western blot analysis. (A) NR2A and NR2B protein levels at P5 and P14 in the developing and denervated PrV. (B) NR2A and NR2B expression on the normal side of the PrV at different ages. The intensity
    Anti Rabbit Nr2a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CRIPT knockdown leads to a selective reduction in the abundance of GluA1 and SAP97. Mixed spinal cord cultures were infected with HSV engineered to express a miRNA targeting CRIPT or a scrambled control. Two days later, lysates were prepared and subjected to Western blottings. No more than six independent experiments were performed for the quantitative image analysis. CRIPT knockdown leads to a reduction in GluA1 and SAP97 abundance and no effect on the abundance of GluA2, GluA4, NR1, NR2A, NR2B, or PSD95. Representative images of Western blottings with actin loading controls are shown and quantification of band intensity in the bar graphs below; *significant difference between groups, p

    Journal: eNeuro

    Article Title: SAP97 Binding Partner CRIPT Promotes Dendrite Growth In Vitro and In Vivo

    doi: 10.1523/ENEURO.0175-17.2017

    Figure Lengend Snippet: CRIPT knockdown leads to a selective reduction in the abundance of GluA1 and SAP97. Mixed spinal cord cultures were infected with HSV engineered to express a miRNA targeting CRIPT or a scrambled control. Two days later, lysates were prepared and subjected to Western blottings. No more than six independent experiments were performed for the quantitative image analysis. CRIPT knockdown leads to a reduction in GluA1 and SAP97 abundance and no effect on the abundance of GluA2, GluA4, NR1, NR2A, NR2B, or PSD95. Representative images of Western blottings with actin loading controls are shown and quantification of band intensity in the bar graphs below; *significant difference between groups, p

    Article Snippet: Acknowledgements: We thank Maria Lim, John Flibotte, and Marco Boccitto for technical assistance and for useful discussion; Yina Dong and David Lynch for the gift of antibodies and plasmids to NR1, NR2A, NR2B, NR2C, and NR2D; Maria Passafaro for the gift of the CRIPT plasmids; and David Miller for the gift of the PPVD ::GFP worms.

    Techniques: Infection, Western Blot

    Immunostaining for cholera toxin β-subunit (CTB) and NMDAR2A in the suprachiasmatic nucleus (SCN) from wild-type (WT) and Shank3 +/– mice. CTB (diaminobenzidine, DAB, left column ) and NMDAR2A ( right column ) did not show obvious differences in the immunoreactivity between groups (WT, top micrographs ; Shank3 +/– , bottom micrographs ). For this and subsequent figures ( Figures 8 , 9 ), scale bar = 50 μm.

    Journal: Frontiers in Neuroscience

    Article Title: Altered Light Sensitivity of Circadian Clock in Shank3+/– Mouse

    doi: 10.3389/fnins.2021.604165

    Figure Lengend Snippet: Immunostaining for cholera toxin β-subunit (CTB) and NMDAR2A in the suprachiasmatic nucleus (SCN) from wild-type (WT) and Shank3 +/– mice. CTB (diaminobenzidine, DAB, left column ) and NMDAR2A ( right column ) did not show obvious differences in the immunoreactivity between groups (WT, top micrographs ; Shank3 +/– , bottom micrographs ). For this and subsequent figures ( Figures 8 , 9 ), scale bar = 50 μm.

    Article Snippet: Primary antibodies were diluted as indicated in 0.1 M PBS containing 1.0% normal serum in 0.3% Triton X-100 [anti-VIP raised in rabbit, CAT 20077, Incstar, 1:2,000 ( ); anti-CTB subunit raised in goat, CAT 703, List Biological Laboratories, 1:2,000 ( ); anti-NMDAR2A (GluN2A) raised in rabbit, CAT ACG 002, Alomone Labs, 1:400 ( ); and anti c-FOS raised in rabbit, CAT SC-52, Santa Cruz, 1:1,000 ( )].

    Techniques: Immunostaining, CtB Assay, Mouse Assay

    Synaptic GluN2B-NMDAR are laterally redistributed in the synaptic area following chem LTP in immature neurons Detection of a single QD (30-Hz acquisition) in our experimental conditions. The high signal-to-noise ratio (SNR) ( > 5) enables the detection and location of the signal with a high pointing accuracy (˜30 nm). The QD fluorescence is quantified on a pseudocolor scale (low: red; high: yellow). Scale bar = 800 nm. A 500-frame stack is obtained while tracking down a single NMDAR/QD complex. On each frame, a single GluN2B- (green) or GluN2A- (blue) QD particle complex is detected and precisely located within synaptic (dark gray) and perisynaptic (320-nm annulus around the synapse; light gray) areas. Those 500 locations are then projected on a single image, providing the successive positions of this receptor/particle complex during the 500-frame stack. Note that GluN2A-NMDAR are more concentrated within the core of the PSD. Scale bar image = 300 nm; synaptic areas = 200 nm. Relative fraction of synaptic and perisynaptic GluN2-QD particles, calculated before and after chem LTP for both GluN2B- (left) and GluN2A-NMDAR (right) ( n = 25 and 20 dendritic fields before and after chem LTP, respectively). Note the significant decrease in the relative synaptic content in GluN2B-NMDAR particles right after chem LTP (Student's t -test, ** P

    Journal: The EMBO Journal

    Article Title: Surface dynamics of GluN2B-NMDA receptors controls plasticity of maturing glutamate synapses

    doi: 10.1002/embj.201386356

    Figure Lengend Snippet: Synaptic GluN2B-NMDAR are laterally redistributed in the synaptic area following chem LTP in immature neurons Detection of a single QD (30-Hz acquisition) in our experimental conditions. The high signal-to-noise ratio (SNR) ( > 5) enables the detection and location of the signal with a high pointing accuracy (˜30 nm). The QD fluorescence is quantified on a pseudocolor scale (low: red; high: yellow). Scale bar = 800 nm. A 500-frame stack is obtained while tracking down a single NMDAR/QD complex. On each frame, a single GluN2B- (green) or GluN2A- (blue) QD particle complex is detected and precisely located within synaptic (dark gray) and perisynaptic (320-nm annulus around the synapse; light gray) areas. Those 500 locations are then projected on a single image, providing the successive positions of this receptor/particle complex during the 500-frame stack. Note that GluN2A-NMDAR are more concentrated within the core of the PSD. Scale bar image = 300 nm; synaptic areas = 200 nm. Relative fraction of synaptic and perisynaptic GluN2-QD particles, calculated before and after chem LTP for both GluN2B- (left) and GluN2A-NMDAR (right) ( n = 25 and 20 dendritic fields before and after chem LTP, respectively). Note the significant decrease in the relative synaptic content in GluN2B-NMDAR particles right after chem LTP (Student's t -test, ** P

    Article Snippet: As previously described (Groc et al , ; Heine et al , ), for the x-link experiments, neurons were co-transfected with GluN1-SEP or GluN2B-SEP and Homer 1c-DsRed and incubated with highly concentrated (1:20) polyclonal antibodies directed against GluN1 (Alomone Labs; epitope corresponding to residues 385–399 of the GluN1 subunit), GluN2B (Alomone Labs; same as above), GluN2A (Alomone Labs; same as above) NMDAR subunits or against GFP (Chemicon).

    Techniques: Fluorescence

    Assessment of NR2A and NR2B expression in the PrV by Western blot analysis. (A) NR2A and NR2B protein levels at P5 and P14 in the developing and denervated PrV. (B) NR2A and NR2B expression on the normal side of the PrV at different ages. The intensity

    Journal: Neuroscience

    Article Title: NMDA RECEPTOR SUBUNIT COMPOSITION IN THE RAT TRIGEMINAL PRINCIPAL NUCLEUS REMAINS CONSTANT DURING POSTNATAL DEVELOPMENT AND FOLLOWING NEONATAL DENERVATION

    doi: 10.1016/j.neuroscience.2011.01.023

    Figure Lengend Snippet: Assessment of NR2A and NR2B expression in the PrV by Western blot analysis. (A) NR2A and NR2B protein levels at P5 and P14 in the developing and denervated PrV. (B) NR2A and NR2B expression on the normal side of the PrV at different ages. The intensity

    Article Snippet: The membranes were incubated in blocking solution (5% milk in TRIS buffered saline with Tween20 (TBS-T) for 1 h at room temperature), then incubated with anti-rabbit NR2A (1:1000 Upstate) or NR2B (1:1000 Alomone Lab) with 5% milk in TBS-T overnight at 4 °C.

    Techniques: Expressing, Western Blot