rabbit polyclonal anti phospho eef2  (Bioss)

Journal: iScience

Article Title: Synaptotagmin-7 deficit causes insulin hypoactivity and contributes to behavioral alterations in mice

doi: 10.1016/j.isci.2025.112354

Figure Lengend Snippet: Signaling pathways involved in the genesis of mood cycling (A) Schematic showing comparative analysis of hippocampal gene expression profiles between manic and depressive Syt7 KO mice. GeneM, comparison between the WT mice and the manic Syt7 KO mice. GeneD, comparison between the WT mice and the depressive Syt7 KO mice. (B and C) Heatmap (B) and distribution (C) of genes differentially expressed between GeneM and GeneD. (D) Expression of representative BD-related genes in manic and depressive Syt7 KO mice. (E) Representative significantly enriched KEGG pathways showing bi-directional alterations in the manic and depressive episodes in Syt7 KO mice. (F and G) Schematic (F) and heatmap (G) showing the comparative analysis of transcripts bi-directionally regulated in drug-induced/Syt7 deficiency-induced manic and depressive mice. GeneR, comparison between the WT mice with and without the Ro25 treatment in the dark phase. GeneB, comparison between the WT mice with and without the BMS-536924 treatment in the light phase. (H) Representative significantly enriched KEGG pathways showing bi-directional alterations between the drug-induced/Syt7-deficiency-induced manic and depressive mice. (I) Mapping of representative KEGG pathways for network analysis. (J) Bi-directional regulation of representative genes within featured KEGG pathways in mice with drug-induced/Syt7-deficiency-induced mania or depression. (K and L) Immunoblots (left) and quantitative analysis (right) of mammalian targets of rapamycin (mTOR) (K) and eukaryotic elongation factor 2 (eEF2) (L) phosphorylation in the hippocampus of WT and Syt7 KO mice. n = 3. Student’s t test; ∗ p < 0.05; ∗∗ p < 0.001; error bars, SEM.

Article Snippet: Rabbit polyclonal anti- p -eEF2 , Cell Signaling , Cat# 2331S; RRID: N/A.

Techniques: Protein-Protein interactions, Gene Expression, Comparison, Expressing, Western Blot, Phospho-proteomics

Journal: iScience

Article Title: Synaptotagmin-7 deficit causes insulin hypoactivity and contributes to behavioral alterations in mice

doi: 10.1016/j.isci.2025.112354

Figure Lengend Snippet: Signaling pathways involved in the genesis of mood cycling (A) Schematic showing comparative analysis of hippocampal gene expression profiles between manic and depressive Syt7 KO mice. GeneM, comparison between the WT mice and the manic Syt7 KO mice. GeneD, comparison between the WT mice and the depressive Syt7 KO mice. (B and C) Heatmap (B) and distribution (C) of genes differentially expressed between GeneM and GeneD. (D) Expression of representative BD-related genes in manic and depressive Syt7 KO mice. (E) Representative significantly enriched KEGG pathways showing bi-directional alterations in the manic and depressive episodes in Syt7 KO mice. (F and G) Schematic (F) and heatmap (G) showing the comparative analysis of transcripts bi-directionally regulated in drug-induced/Syt7 deficiency-induced manic and depressive mice. GeneR, comparison between the WT mice with and without the Ro25 treatment in the dark phase. GeneB, comparison between the WT mice with and without the BMS-536924 treatment in the light phase. (H) Representative significantly enriched KEGG pathways showing bi-directional alterations between the drug-induced/Syt7-deficiency-induced manic and depressive mice. (I) Mapping of representative KEGG pathways for network analysis. (J) Bi-directional regulation of representative genes within featured KEGG pathways in mice with drug-induced/Syt7-deficiency-induced mania or depression. (K and L) Immunoblots (left) and quantitative analysis (right) of mammalian targets of rapamycin (mTOR) (K) and eukaryotic elongation factor 2 (eEF2) (L) phosphorylation in the hippocampus of WT and Syt7 KO mice. n = 3. Student’s t test; ∗ p < 0.05; ∗∗ p < 0.001; error bars, SEM.

Article Snippet: Primary antibodies were as follows: mouse monoclonal anti-Actin antibody (1:5000, Abcam, #ab6276), rabbit polyclonal anti-mTOR antibody (1:500, Cell Signaling, #2972), rabbit polyclonal anti- p -mTOR antibody (1:500, Cell Signaling, #2971), rabbit polyclonal anti-eEF2 antibody (1:1000, Cell Signaling, #2332S), rabbit polyclonal anti- p -eEF2 antibody (1:1000, Cell Signaling, #2331S).

Techniques: Protein-Protein interactions, Gene Expression, Comparison, Expressing, Western Blot, Phospho-proteomics

Journal: Cell reports

Article Title: Convergence of distinct signaling pathways on synaptic scaling to trigger rapid antidepressant action

doi: 10.1016/j.celrep.2021.109918

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-phospho-eEF2 , Cell Signaling Technology , Cat# 2331; RRID:AB_10015204.

Techniques: Recombinant, Software

Journal: iScience

Article Title: Secretory mouse quiescin sulfhydryl oxidase 1 aggregates defected human and mouse spermatozoa in vitro and in vivo

doi: 10.1016/j.isci.2021.103167

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-EEF2 , Abcam , #ab40812; RRID: AB_732082.

Techniques: Plasmid Preparation, Recombinant, Purification, Northern Blot, Software

Journal: BMC Biochemistry

Article Title: ADP-Ribosylargininyl reaction of cholix toxin is mediated through diffusible intermediates

doi: 10.1186/s12858-014-0026-1

Figure Lengend Snippet: Intramolecular transfer of ADP-ribose to multiple arginines in the auto-reaction. (A) Mixing marked enzymes experiments. Active Ig-CTc was mixed with inactive mutant Y493A (top panel); active full-length cholix toxin, CXT (WT), was mixed with mutant Y493A (middle panel); inactive mutant CXT(E581A) was mixed with active CTc (bottom panel) in auto-ADP-ribosylation assays. Molar concentrations shown on the top of each blot are the concentrations of active enzyme. The left blots (SA) were detected by IRDye800CW-SA; the same blots were re-probed with polyclonal rabbit anti-CTc antibody shown on right (IB). (B) Neutral hydroxylamine assays. Auto-ADP-ribosylation reaction was carried out with periplasmic fractions of E. coli lysate expressing wild type or mutant CTc. The products were subjected to 0.5 M NH 2 OH (pH7.5) or 0.5 M NaCl treatment at 37°C for 2 hours. The blot shown is representative of multiple experiments. (C-E) Single amino acid substitution mutation studies. A set of representative data from the detection of biotin signals on wild type or different mutants was shown in (C) . The fluorescence intensity of SA (IRDy-SA) was normalized with the Coomassie Blue intensity (CB) to obtain the IRDy-SA/CB ratio as a semi-quantitative auto-ADP-ribosylation measurement of each enzyme. Data were summarized from four sets of experiments (D) . Asterisk indicates significant reduction of auto-ADP-ribosylation activity. NAD + glycohydrolase activities of wild type and mutant enzymes are shown in (E) .

Article Snippet: Wild type and mutant CTc or eEF2 in each transferred membrane were exposed to rabbit polyclonal anti-CTc antibody, rabbit polyclonal anti-eEF2 antibody (Abcam), or mouse anti-flag (M2) monoclonal antibody (Sigma) followed by different secondary antibodies.

Techniques: Mutagenesis, Expressing, Fluorescence, Activity Assay

Journal: BMC Biochemistry

Article Title: ADP-Ribosylargininyl reaction of cholix toxin is mediated through diffusible intermediates

doi: 10.1186/s12858-014-0026-1

Figure Lengend Snippet: ADP-ribosylation of endogenous versus exogenous substrates. (A-B) Cholix toxin catalytic fragments were pre-incubated with or without 50 μM NAD + at 37°C for 1 hour. Free NAD + was removed by gel filtration chromatography. The recovered enzymes were quantified. Equal concentrations of auto-ADP-ribosylated CTc (pre-incubated with NAD + ) or control (non-auto-ADP-ribosylated CTc, processed through auto-ADP-ribosylation reaction without NAD + ) were used in the NAD + glycohydrolase activity assays and ADP-ribosylation of eEF2 in 293 lysate. (C) His-tagged oligo-L-arginine or oligo-L-asparagine peptides were incubated with purified recombinant wild type CTc, catalytically defective mutant (Y493A) or catalytically active mutant (E579Q). The samples were analyzed by a 96-well plate based ADP-ribosylation assay. Data shown are composite from two experiments with triplicates within-plate replicates. (D) Various concentrations of catalytic fragments of cholix toxin or exotoxin A (PEA) was incubated with CHO or Re1.22c cell lysate at 37°C for 1 hr. The biotin signals on the ADP-ribosylated eEF2 and auto-ADP-ribosylated enzymes were detected by IRDye800CW-SA shown on the top panel. The same blot was stripped and re-probed with anti-CTc and anti-eEF2 antibodies shown in the middle panel. The bottom panel shows the Coomassie Blue stained gel for protein loading control. (E) To detect auto-ADP-ribosylation at the presence of exogenous substrates, excess amount of CTc (6 μM) was incubated with purified flag-tagged wild type eEF2 (0.2 μM) or flag-tagged eEF2 (H715R) mutant (0.2 μM) at the presence of 50 μM biotinyl-NAD + for various periods of time.

Article Snippet: Wild type and mutant CTc or eEF2 in each transferred membrane were exposed to rabbit polyclonal anti-CTc antibody, rabbit polyclonal anti-eEF2 antibody (Abcam), or mouse anti-flag (M2) monoclonal antibody (Sigma) followed by different secondary antibodies.

Techniques: Incubation, Filtration, Chromatography, Control, Activity Assay, Purification, Recombinant, Mutagenesis, Staining