pe labelled mouse anti human trka  (R&D Systems)

Journal: Biomedicines

Article Title: A Possible Role for Nerve Growth Factor and Its Receptors in Human Sperm Pathology.

doi: 10.3390/biomedicines11123345

Figure Lengend Snippet: Figure 2. Nerve growth factor (NGF) receptors in the ejaculated sperm, epididymis and testis of fertile men. (A) The bar graph shows the mean and standard deviation of the fold increase in TrKA (slanted-line bar graph) and p75NTR (checkerboard bar graph) messenger RNA (mRNA) levels in fertile men (n = 7). (B,C) The plots (blue gate) display the flow cytometric analysis (representative of

Article Snippet: After washing three times in PBS supplemented with BSA, sperm were incubated at 4 ◦C for 1 h in PBS/BSA containing FITC-labelled rabbit anti-human p75NTR extracellular domain (ANT-007-F, Alomone Labs, Jerusalem, Israel) (10 mL/sample) and PE-labelled mouse anti-human TrKA (FAB1751P, R&D Systems, Minneapolis, MN, USA) (10 mL/sample).

Techniques: Standard Deviation

Journal: Biomedicines

Article Title: A Possible Role for Nerve Growth Factor and Its Receptors in Human Sperm Pathology.

doi: 10.3390/biomedicines11123345

Figure Lengend Snippet: Figure 3. Nerve growth factor (NGF) receptors in altered sperm from men with infertility. (A–D) Fluorescence immunostaining of human sperm from men with infertility (varicocele (V) or uro- genital infections (UGIs)) incubated with the anti-TrKA (A,B) and anti-p75NTR (C,D) antibodies. In (A)

Article Snippet: After washing three times in PBS supplemented with BSA, sperm were incubated at 4 ◦C for 1 h in PBS/BSA containing FITC-labelled rabbit anti-human p75NTR extracellular domain (ANT-007-F, Alomone Labs, Jerusalem, Israel) (10 mL/sample) and PE-labelled mouse anti-human TrKA (FAB1751P, R&D Systems, Minneapolis, MN, USA) (10 mL/sample).

Techniques: Fluorescence, Immunostaining, Incubation

Journal: PLoS ONE

Article Title: Chronic Nerve Growth Factor Exposure Increases Apoptosis in a Model of In Vitro Induced Conjunctival Myofibroblasts

doi: 10.1371/journal.pone.0047316

Figure Lengend Snippet: Confluent myoFB were exposed to NGF at the indicated doses/times and evaluated for biochemical and molecular changes. (A) αSMA protein expression in both myoFB and FB, as detected by cell surface ELISA (optic density) and flow cytometry (fluorescence intensity). (B–C) Fluorescent histograms showing a different pattern of trkA NGFR and p75 NTR expression in acute and chronic 100 ng/mL NGF exposed myoFB, as compared to untreated myoFB (black line). Mean Fluorescent Intensity (MFI) plots are shown inside fluorescent histograms (iso stands for isotype-matched control FI; 7.05 for trkA NGFR and 8.39 for p75 NTR ). (D) Morphological distribution of trkA NGFR /p75 NTR in untreated, acute and chronic 100 ng/mL NGF treated myoFB. (E–F) Target gene expression specific for trkA NGFR /p75 NTR after acute and for αSMA/p75 NTR after chronic exposure to increasing NGF (p<.05).

Article Snippet: Cells were incubated with mouse anti-human αSMA antibodies (sc-130616) followed by species-specific APC-conjugated anti-Mouse IgG antibodies; or probed with a PE-conjugated anti-human trkA NGFR antibodies (sc-118) and FC-conjugated anti-human p75 NTR antibodies (sc-81612) mixture (Santa Cruz Biotech., Santa Cruz, CA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Fluorescence

Journal: PLoS ONE

Article Title: Chronic Nerve Growth Factor Exposure Increases Apoptosis in a Model of In Vitro Induced Conjunctival Myofibroblasts

doi: 10.1371/journal.pone.0047316

Figure Lengend Snippet: (A) Pictures depicting TUNEL reactivity in p75 NTR positive cells exposed to chronic 100 ng/mL NGF, as compared to TGFβ1 exposed cells. Changes in morphology, such as having rounded cytoplasm and markedly condensed chromatin/nuclear fragmentation, are clear visible in p75 NTR -bearing myoFB. A few TUNEL-positive myoFB were also quantified in untreated cultures (not shown). (B) Overlays and single staining specific for p75 NTR and cleaved caspase3 are visible in both acute and chronic 100 ng/mL NGF exposed myoFB, as compared to untreated myoFB. (C–D) p75 NTR positive sorted cells analysed for active caspase3 expression and related statistical analysis of MFI. Note the specific reduction upon pretreated with trkA NGFR and/or p75 NTR chemical inhibitors.

Article Snippet: Cells were incubated with mouse anti-human αSMA antibodies (sc-130616) followed by species-specific APC-conjugated anti-Mouse IgG antibodies; or probed with a PE-conjugated anti-human trkA NGFR antibodies (sc-118) and FC-conjugated anti-human p75 NTR antibodies (sc-81612) mixture (Santa Cruz Biotech., Santa Cruz, CA).

Techniques: TUNEL Assay, Staining, Expressing

Journal: PLoS ONE

Article Title: Chronic Nerve Growth Factor Exposure Increases Apoptosis in a Model of In Vitro Induced Conjunctival Myofibroblasts

doi: 10.1371/journal.pone.0047316

Figure Lengend Snippet: (A) A flow chart schematizing the siRNA experiments. (B) Confocal images depicting GFP positive cells (image) and p75 NTR -GFP double-positive cells (insert over the image), re-plated 5 hrs after electroporation. (C–D) Representative histogram for GFP specific staining, in comparison to control siRNA (control cells), showing a decreased p75 NTR positive expression and the related p75 NTR target gene deregulation. (E) Mean Fluorescence Intensity (MFI) specific for caspACE-VAD comparing 100 ng/mL NGF and 10 ng/mL TGFβ1 effects on control-siRNA, p75 NTR siRNA and double trkA NGFR /p75 NTR siRNA treated cells. A 37.5%-decrease (*) and a 50%-decrease (**) of apoptotic signal were detected respectively in p75 NTR -siRNA and p75 NTR /trkA NGFR -siRNA (p<.05).

Article Snippet: Cells were incubated with mouse anti-human αSMA antibodies (sc-130616) followed by species-specific APC-conjugated anti-Mouse IgG antibodies; or probed with a PE-conjugated anti-human trkA NGFR antibodies (sc-118) and FC-conjugated anti-human p75 NTR antibodies (sc-81612) mixture (Santa Cruz Biotech., Santa Cruz, CA).

Techniques: Electroporation, Staining, Expressing, Fluorescence

Journal: Frontiers in Urology

Article Title: The Role of NGF and Its Receptor TrKA in Patients With Erectile Dysfunction

doi: 10.3389/fruro.2022.860612

Figure Lengend Snippet: FIGURE 1 | (A) Medium NGF levels in PBMC-treated (20 mg/ml or 20 pg/ml of ALP) or untreated cells. Description of what is contained in the first panel; (B) TrKA and (C) p75NTR Facs analysis of PBMC-treated (20 mg/ml or 20 pg/ml of ALP) or untreated cells. CTR, control; EtOH, alprostadil in ethanol; ALP, alprostadil.

Article Snippet: The following fluorochrome-labeled mAbs were used to identify TrKA and p75NTR receptors onWBCs: FITClabeled rabbit anti-human p75NTR extracellular domain (#ANT007-F, Alomone Labs) and PE-labeled mouse anti-human TrKA (FAB1751P, R&D Systems).

Techniques: Control

Journal: Frontiers in Urology

Article Title: The Role of NGF and Its Receptor TrKA in Patients With Erectile Dysfunction

doi: 10.3389/fruro.2022.860612

Figure Lengend Snippet: FIGURE 4 | (A) Colorimetric analysis of total thiol in plasma of ED patients. Cubital vein of healthy subjects (CV-CTRL), cubital vein (CV) or corpus cavernosum (CC) of ED patients after alprostadil (ALP) injection (CV+ALP), and CC after ALP (CC+ALP). Horizontal bars indicate mean values. ***p < 0.0001. Comparisons between the groups were made using one-way ANOVA followed by Bonferroni multiple comparison tests. (B) Correlation between TrKA+ cells and thiol levels in CC-ED. r, Pearson index; r2, coefficient of determination.

Article Snippet: The following fluorochrome-labeled mAbs were used to identify TrKA and p75NTR receptors onWBCs: FITClabeled rabbit anti-human p75NTR extracellular domain (#ANT007-F, Alomone Labs) and PE-labeled mouse anti-human TrKA (FAB1751P, R&D Systems).

Techniques: Clinical Proteomics, Injection, Comparison

Journal: Frontiers in Urology

Article Title: The Role of NGF and Its Receptor TrKA in Patients With Erectile Dysfunction

doi: 10.3389/fruro.2022.860612

Figure Lengend Snippet: FIGURE 3 | FACS analysis of TrKA in blood withdrawals of ED patients. (A) Percentage of TrKA+ cells; (B) MFI: mean fluorescent intensity. Cubital vein of healthy subjects (CV-CTRL), cubital vein (CV) or corpus cavernosum (CC) of ED patients after Alprostadil (ALP) injection (CV+ALP), and CC after ALP (CC+ALP). Horizontal bars indicate mean values. ***p < 0.0001; **p < 0.001; *p < 0.01. Comparisons between the groups were made using one-way ANOVA followed by Bonferroni multiple comparison tests. CV-CTRL.

Article Snippet: The following fluorochrome-labeled mAbs were used to identify TrKA and p75NTR receptors onWBCs: FITClabeled rabbit anti-human p75NTR extracellular domain (#ANT007-F, Alomone Labs) and PE-labeled mouse anti-human TrKA (FAB1751P, R&D Systems).

Techniques: Injection, Comparison