well rabbit novus biologicals nbp1 48336 ldh a antibody blocking peptide elisa detection n a n a novus biologicals nbp1 48336pep ldh b elisa detection  (Novus Biologicals)

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) TRPV3 immunostaining (green) in mouse primary somatosensory (S1) cortex, striatum, thalamus, and hippocampus, with DAPI counterstain (blue). Top row left: Postnatal day (P)7; bottom row left: P14; bottom row right: P21. Top row right: Sections incubated with TRPV3 antibody plus TRPV3 blocking peptide in S1 cortex, striatum, thalamus, and hippocampus (green) with DAPI counterstain (blue). Remaining TRPV3 staining is shown in green with DAPI in blue. ( B ) TRPV3 and TRPV4 immunostaining in S1 cortex, striatum, thalamus, and hippocampus at P14. Top row left: TRPV3 (green) and TRPV4 (brick red). Top row right: TRPV3 (green) and TRPV4 (brick red) with DAPI counterstain (blue). Bottom row left: TRPV4 immunostaining in S1BF (inset from top row left) at 10x. Bottom row right: TRPV3 immunostaining in S1BF (inset from top row left) at 10x.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Immunostaining, Incubation, Blocking Assay, Staining

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording whole-cell TRPV3 currents at 30°C (black), 36°C (gray), and 39°C (red) in cortical excitatory pyramidal neurons (PNs) with bath application of camphor (5 mM), a TPRV3 agonist. ( B ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black), 36°C (gray) and 39°C (red) in wild-type (WT) mice: 11 cells from four mice. ( C ) Scatter dot plots of the current density-voltage measurements. ( D ) Current density-voltage (I–V) relationship of TRPV3 currents at 30°C (black) in the presence of camphor (5 mM), a TPRV3 agonist, or camphor (5 mM) + TRPV3 blocker (Forsythoside B, 50 µM) (blue). ( E ) Same as ( D ) but for 36°C. ( F ) Same as ( D ) but for 39°C. ( G ) Current density-voltage (I–V) plot showing the net TRPV3 current (opener – (opener+blocker) condition). In B - F , statistical significance was assessed using a two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques:

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potential and spiking in an excitatory cortical pyramidal neuron (PN) with an intracellular blocker of TRPV3 channels (Forsythoside B, 50 µM) (left) or TRPV4 channels (RN1734, 10 µM) (right) at just-subthreshold V m at 30°C, 36°C, and 39°C in mouse S1 cortex. ( B ) Percentages of cell types obtained from experiment in A . ( C ) Evoked spikes in L2/3 cortical PNs during temperature elevations to 30°C, 36°C, and 39°C under three conditions: no blockers, TRPV3 blocker (Forsythoside B, 50 µM), or TRPV4 blocker (RN1734, 10 µM). ( D ) Correlation between post-synaptic potential (PSP) peak and spike threshold (ST). r =Pearson correlation coefficient with Deming linear regression. ( E ) Same as ( C ) for the L4-evoked late PSP peak. ( F ) Same as ( C ) for input resistance (R in ). Each data point in C – F represents an individual cell. Data were collected from 26 cells in 7 animals for the TRPV3 blocker, 24 cells in 6 animals for the TRPV4 blocker, and 37 cells in 14 animals for the no-block condition. Mean ± SEM is shown in C , E , and F . Statistical significance was assessed using one- or two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). In D , correlations were evaluated using Pearson’s r with Deming linear regression.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Blocking Assay

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) Setup for recording L4-evoked post-synaptic potentials and spiking in excitatory cortical pyramidal neurons (PNs) at just-subthreshold Vm at 30°C, 36°C, and 39°C in mouse S1 cortex of wild-type ( Trpv3 +/+ ) and Trpv3 knockout ( Trpv3 -/- ) mice. ( B ) Depolarization required to reach spike threshold (ST) in Trpv3 +/+ and Trpv3 -/- mice at 30°C, 36°C, and 39°C. ( C ) Same as ( B ) but for input resistance (R in ). ( D ) Same as ( B ) but for number of spikes. ( E ) Same as ( B ) but for post-synaptic potential (PSP). ( F ) Setup for recording mouse body temperature (T b ) at room temperature and during fever-range and higher, using an implanted transponder for non-invasive measurement, with exposure to infrared light. ( G ) Time to loss of postural control (LPC), defined as collapse and failure to maintain upright posture, in wild-type ( Trpv3 +/+ ), heterozygous ( Trpv3 +/- ), and Trpv3 knockout ( Trpv3 -/- ) mice. ( H ) Same as in ( G ) but showing T b at seizure onset. ( I ) Same as in ( G ) but for the time from LPC to seizure onset. Each data point in B – E represents an individual cell (three animals per genotype). Statistical significance was assessed using two-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05). Each data point in G – I represents an individual animal. Statistical significance was assessed using one-way repeated-measures ANOVA with Tukey’s or Sidak post-hoc test ( α =0.05).

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Knock-Out, Control

Journal: eLife

Article Title: TRPV3 channel activity helps cortical neurons stay active during fever

doi: 10.7554/eLife.102412

Figure Lengend Snippet: ( A ) In cortical L2/3 pyramidal neurons (PNs) with synaptically evoked spiking, gradual increases in brain slice temperature from 30 °C to 36°C to 39°C result in four possible outcomes: neurons remain inactive, continue spiking (STAY), stop spiking, or initiate spiking. To spike, PNs must reach the spike threshold (ST), defined as the minimal V m that elicits an action potential, which requires sufficient depolarization via the post-synaptic potential (PSP). STAY neurons consistently reach ST and continue spiking. Neurons that stop spiking fall below the level of depolarization required to reach ST, while neurons that initiate spiking achieve sufficient depolarization to newly reach ST. ( B ) STAY neurons may contain unique ion channels, such as TRPV3. TRPV3 channels are highly permeable to Ca² + ions, and Ca² + influx contributes to PN depolarization. The presence of TRPV3 facilitates greater ion entry, enabling depolarization sufficient to reach ST and sustain spiking, whereas its absence reduces depolarization to levels insufficient for spiking.

Article Snippet: Slides were blocked in 10% normal goat serum for 1 hr at room temperature, then incubated overnight at 4°C with anti-TRPV3-Biotin antibody (#ACC-033-B), TRPV3 blocking peptide (#BLP-CC033), and/or anti-TRPV4 antibody (#ACC-034) (Alomone Labs, Israel).

Techniques: Slice Preparation

Journal: Cardio-oncology

Article Title: Cardiomyocyte overexpression of microRNA-210 mitigates apoptotic cell death induced by doxorubicin

doi: 10.1186/s40959-025-00429-z

Figure Lengend Snippet: miR-210 reduces DOX-induced apoptotic cell death in AC-16 cardiomyocytes. A Overexpressing miR-210 (OE) significantly attenuated caspase-3 activity, while ( B ) knocking down miR-210 (KD) significantly increased caspase-3 activity. Absorbance was measured at 405 nm. Caspase-3 activity was corrected for total protein content in each sample measured by β- Actin. All data are expressed as mean ± S.D. from three technical replicates for each of the four biological replicates ( n = 4) belonging to each experimental group. **** p < 0.0001. DOX: doxorubicin; OE: miR-210 overexpression; KD: miR-210 knockdown; S.D.: standard deviation. Notation: “ + ” indicates treatment applied; “–” indicates absence of treatment. For DOX, “ + ” = DOX-treated, “–” = vehicle. For miR-210 OE, “ + ” = miR-210 OE, “–” = empty vector. For miR-210 KD, “ + ” = miR-210 knockdown, “–” = empty vector

Article Snippet: β-Actin antibody blocking peptide , ELISA detection , N/A , N/A , Cell Signalling Technology , 1025.

Techniques: Activity Assay, Over Expression, Knockdown, Standard Deviation, Plasmid Preparation

Journal: Cardio-oncology

Article Title: Cardiomyocyte overexpression of microRNA-210 mitigates apoptotic cell death induced by doxorubicin

doi: 10.1186/s40959-025-00429-z

Figure Lengend Snippet: miR-210 overexpression increases phosphorylation at serine 9 residue (Ser9) of glycogen synthase kinase-3β (GSK-3β)in DOX-treated AC-16 cardiomyocytes. A Phosphorylation at Ser9 of GSK-3β is increased with miR-210 OE vector as compared to EV after 24-h DOX treatment. B No significant difference in phosphorylation at Ser9 of GSK-3β is observed with miR-210 KD vector as compared to EV after 24-h DOX treatment. No change in protein expression levels of GSK-3βwere observed in AC-16 cardiomyocytes transfected with an overexpression (OE) vector ( C ) nor a knockdown (KD) vector ( D ) of miR-210 across all treatment arms. Protein expression of phosphorylation at Ser9 of GSK-3β was further confirmed by western blot under the condition of ( E ) miR-210 OE and ( F ) mir-210 KD. Protein expression of GSK-3β was corrected for total protein content in each sample measured by β- Actin. Absorbance was measured at 405 nm. All data are expressed as mean ± S.D. All data for ELISA analyses ( A - D ) was run from three technical replicates for each of the four biological replicates ( n = 4) belonging to each experimental group. Panel G display exemplary western blot. Samples for was performed on three random selected samples from each group presented in panel A-D to allow comparison on the same gel.** p < 0.01; **** p < 0.0001. DOX: doxorubicin; OE: miR-210 overexpression; KD: miR-210 knockdown; S.D.: standard deviation. Notation: “ + ” indicates treatment applied; “–” indicates absence of treatment. For DOX, “ + ” = DOX-treated, “–” = vehicle. For miR-210 OE, “ + ” = miR-210 OE, “–” = empty vector. For miR-210 KD, “ + ” = miR-210 knockdown, “–” = empty vector

Article Snippet: β-Actin antibody blocking peptide , ELISA detection , N/A , N/A , Cell Signalling Technology , 1025.

Techniques: Over Expression, Phospho-proteomics, Residue, Plasmid Preparation, Expressing, Transfection, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison, Standard Deviation

Journal: Cardio-oncology

Article Title: Cardiomyocyte overexpression of microRNA-210 mitigates apoptotic cell death induced by doxorubicin

doi: 10.1186/s40959-025-00429-z

Figure Lengend Snippet: miR-210 affects phosphorylation of Akt at serine 473 (Ser473) in DOX-treated AC-16 cardiomyocytes. No change in protein expression levels of Akt were observed in AC-16 cardiomyocytes transfected with either ( A ) the overexpression (OE) vector of miR-210 or the knock down vector (KD) ( B ). C Phosphorylation at Ser473 of Akt is increased with miR-210 OE vector as compared to EV after 24-h DOX treatment. D A significantly lower degree of phosphorylation at Ser473 of Akt is observed with miR-210 KD vector as compared to empty vector (EV) after 24-h DOX treatment. All data for ELISA analyses was run from three technical replicates for each of the four biological replicates ( n = 4) belonging to each experimental group. Expression levels of phosphorylation at Ser473 of Akt was further confirmed by western blot under the condition of ( E ) the overexpression miR-210 OE and ( F ) mir-210 KD. Absorbance was measured at 405 nm. Protein expression of Akt was corrected for total protein content in each sample measured by β- Actin that also served as control for expression of GSK-3β protein loading. Panel G display exemplary western blot. Samples for was performed on three random selected samples from each group presented in panel A-D to allow comparison on the same gel. * p < 0.05; ** p < 0.01; **** p < 0.0001. DOX: doxorubicin; OE: miR-210 overexpression; KD: miR-210 knockdown; S.D.: standard deviation. -/+ to each group for DOX denotes if the condition of the cells were with DOX (+) or with vehicle treatment (-). For miR-210 OE -/+ denotes if cells where treated by miR-210 OE (+) or denotes the treatment were EV (-). For miR-210 KD -/+ denotes if cells were treated by miR-210 KD (+) or denotes the treatment were EV (-)

Article Snippet: β-Actin antibody blocking peptide , ELISA detection , N/A , N/A , Cell Signalling Technology , 1025.

Techniques: Phospho-proteomics, Expressing, Transfection, Over Expression, Plasmid Preparation, Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot, Control, Comparison, Standard Deviation, IF-cells