rabbit polyclonal anti p2x 7 receptor antibody  (Alomone Labs)

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: P2X 7 immunoreactivity in mouse normal skin (A), papilloma (C), and skin cancer tissues (E) (Experiment-1); B, D, F are parallel cross sections, respectively, stained by H&E . G . Analysis of P2X 7 immunoreactivity compared among paired histologically normal and cancerous tissues. Bars are means (± SD) of levels in tissues of five mice. H . P2X 7 protein assays in mouse normal and cancer skin tissues. Lysates fractionated by gel electrophoresis were immunoblotted with the anti P2X 7 antibody and membranes were reprobed with the anti GAPDH antibody. Insert in H shows Western immunoblot of lysates of histologically normal and cancerous tissues obtained from the same animal. Similar results were obtained in tissues of two additional mice. Bars show means (± SD) of densitometry results of the P2X 7 -specific 75 KDa bands in tissues of three mice. I . P2X 7 mRNA levels (relative to GAPDH mRNA) (means ± SD) in histologically normal and cancerous tissues obtained from three animals. In G-I data in the normal tissues were normalized in each case to an arbitrary value of 10. * – p < 0.01. AU – arbitrary units.

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques: Staining, Nucleic Acid Electrophoresis, Western Blot

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: Effects in mice in-vivo of local treatment with BzATP on skin apoptosis (Experiment-3) . Animals (n = 5) were treated for 16 weeks either with BzATP, applied locally twice a week on the shaved dorsal skin ( A, B ) or with the vehicle (Control, n = 5, C, D ). At the end of the experiment animals were euthanized and strips were obtained from each animal dorsal skin areas for H&E ( B, D ) and P2X 7 /TUNEL co-staining ( E-L ). Arrows in J show increased TUNEL staining co-localizing with P2X 7 immunoreactivity in epidermal cells of the basal/parabasal layers (horizontal arrow) and of epidermal hair shaft cells (vertical arrow). Assays were repeated 3–5 times with similar trends.

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques: In Vivo, TUNEL Assay, Staining

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: Effects of treatments with BzATP on P2X 7 expression and apoptosis in DMBA/TPA – induced skin papillomas (A-D) and cancers (E-H) (Experiment-1) . Assays were repeated 4 times with similar trends. C, D, G, H are parallel cross sections to A, B, E, F , respectively.

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: Effects of BzATP on apoptosis in cultured mouse keratinocytes . In all experiments levels of apoptosis were normalized to an arbitrary value of 2 in control cells. AU – arbitrary units. A . BzATP dose-response effect in cultured mouse normal (wild-type, C57Bl) keratinocytes (means ± SD, n = 3). Cells were treated with one of the indicated concentrations of BzATP for 8 hours. B . Cultured mouse normal keratinocytes (wild-type, C57Bl) were pre-treated with 100 μM anti-sense P2X 7 oligonucleotides (ASO) or random-control P2X 7 oligonucleotides (RCO) for 14 hours followed by 8 hours treatment with 100 μM BzATP. Control – cells treated with the vehicle of the ASO. Values are means (± SD) of 3 experiments for each condition. Insert in B is Western immunoblot with anti-P2X 7 antibody of lysates of cells treated with ASO or RCO (n = 2). C . BzATP time-response effect in cultured mouse normal keratinocytes (wild-type, C57Bl; filled triangles), or in keratinocytes obtained from P2X 7 -deficient (P2X 7 -/-Pfizer [empty circles] or P2X 7 -/-GSK [filled circles]) mice (both in the C57Bl background). Values are means (± SD) of 3 experiments for each condition. Changes in apoptosis in A and B were determined in terms of solubilized DNA; changes in apoptosis in C were determined using cell-death detection ELISA.

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques: Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: Effects of anti-sense P2X 7 oligonucleotides on BzATP-induced increase in cytosolic calcium (ΔCa 2+ i , A) and on the influx of ethidium bromide (Eth-Br, B) . Cultured mouse wild-type normal keratinocytes were pre-treated with 100 μM anti-sense P2X 7 oligonucleotides (ASO) or random-control P2X 7 oligonucleotides (RCO) for 14 hours followed by 8 hours treatment with 100 μM BzATP. Control – cells treated with the vehicle of the ASO. Levels of ΔCa 2+ i ( A ) and of Eth-Br fluorescence ( B ) were determined as in Fig. 12. The experiments were repeated twice with similar trends.

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques: Cell Culture, Fluorescence

Journal: BMC Cancer

Article Title: Activation of P2X 7 -mediated apoptosis Inhibits DMBA/TPA-induced formation of skin papillomas and cancer in mice

doi: 10.1186/1471-2407-9-114

Figure Lengend Snippet: Effects of pre-treatment with anti-sense P2X 7 oligonucleotides (ASO) or random-control P2X 7 oligonucleotides (RCO) (both at 100 μM for 14 hours), and of treatments with BzATP (100 μM, 8 hours) on [ 3 H]thymidine incorporation in mouse wild-type normal keratinocytes (values are means ± SD, n = 4) .

Article Snippet: Primary antibodies for the immunostaining and Western blots were as follows: Rabbit polyclonal anti-P2X 7 receptor antibody, which recognizes the functional full length P2X 7 receptor [ ] was from Alomone Laboratories (Jerusalem, Israel); rabbit anti-GAPDH antibody was from BD Transduction Laboratories (Lexington, KY).

Techniques:

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: ATP evokes Ca 2+ responses and CXCL5 secretion via P2X 4 receptor activation in human monocyte-derived macrophages

doi: 10.4049/jimmunol.1700965

Figure Lengend Snippet: A) Bright field image to study morphology of GM-MDM cells resembling classical ‘fried-egg’ characteristics. Scale bar represents 10 μm. B) Flow cytometer analysis: FSC vs. SSC plot with P2 population indicating MDM population (left panel) and histogram to quantify CD14+ GM-MDM cells (Red: IgG Isotype control, Green: CD14-PE; right panel). C) Level of mRNA expression of P2X and P2Y receptor genes as quantified by qRT-PCR (N=5 donors). mRNA transcript found above Ct 35 is considered absent. D) Distribution and expression of intracellular P2X and P2Y receptor proteins in GM-MDM cells as visualized under confocal microscopy. Secondary only control with either goat AF488 or Rabbit AF488 are included on the left. Scale bar represents 10 μm. Images shown in A, B and D are representative of 3 donors.

Article Snippet: Cells were blocked with 1% BSA (30min, RT) and incubated overnight at 4°C with primary antibodies (rabbit polyclonal P2X 4,7 and P2Y 11,13 (Alomone, Israel), goat polyclonal P2X 1,5 and P2Y 12 , and rabbit polyclonal P2Y 1,2 (Santa-Cruz Biotechnology, USA)).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Confocal Microscopy

Journal: PLoS ONE

Article Title: Effect of naringin on gp120-induced injury mediated by P2X 7 receptors in rat primary cultured microglia

doi: 10.1371/journal.pone.0183688

Figure Lengend Snippet: A, Relative expression of P2X 7 mRNA detected by qPCR. B, SDS-PAGE electrophoresis of P2X 7 receptor protein. C, Relative expression of P2X 7 receptor protein; ** P < 0.01, versus Ctrl group; n = 5.

Article Snippet: The primary antibodies were as follows: rabbit polyclonal anti-P2X 7 (Alomone, Israel; 1:400 dilution), and β-actin (Beijing Zhongshan Biotech Co., 1:800 dilution).

Techniques: Expressing, SDS Page, Electrophoresis

Journal: PLoS ONE

Article Title: Effect of naringin on gp120-induced injury mediated by P2X 7 receptors in rat primary cultured microglia

doi: 10.1371/journal.pone.0183688

Figure Lengend Snippet: A, SDS-PAGE electrophoresis of P2X 7 receptor proteins in primary cultured microglia. B, Relative expression level of P2X 7 receptor protein in primary cultured microglia; ** P < 0.01, versus Ctrl group; ## P < 0.01, versus gp120 group; n = 5.

Article Snippet: The primary antibodies were as follows: rabbit polyclonal anti-P2X 7 (Alomone, Israel; 1:400 dilution), and β-actin (Beijing Zhongshan Biotech Co., 1:800 dilution).

Techniques: SDS Page, Electrophoresis, Cell Culture, Expressing

Journal: PLoS ONE

Article Title: Effect of naringin on gp120-induced injury mediated by P2X 7 receptors in rat primary cultured microglia

doi: 10.1371/journal.pone.0183688

Figure Lengend Snippet: A, Typical fluorescence image of primary cultured microglia; green signal represents CD11b staining with FITC; red signal indicates P2X 7 staining with TRITC; Merge represents the P2X 7 and CD11b double staining image; arrows represent microglia with co-expression of P2X 7 receptor and CD11b. B, Co-expression values of CD11b and P2X 7 receptor in each group; ** P < 0.01, versus Ctrl group; ## P < 0.01, versus gp120 group; n = 3.

Article Snippet: The primary antibodies were as follows: rabbit polyclonal anti-P2X 7 (Alomone, Israel; 1:400 dilution), and β-actin (Beijing Zhongshan Biotech Co., 1:800 dilution).

Techniques: Fluorescence, Cell Culture, Staining, Double Staining, Expressing

Journal: Purinergic Signalling

Article Title: Distribution of purinergic P2X receptors in the equine digit, cervical spinal cord and dorsal root ganglia

doi: 10.1007/s11302-013-9356-5

Figure Lengend Snippet: Primer sequences (forward/reverse) for equine P2X 1−7 cDNAs

Article Snippet: Membranes were blocked in 5 % skimmed dry milk in phosphate-buffered saline for 1 h at room temperature and incubated overnight at 4 °C with the primary antibody in 0.1 % Tween blocking solution, (1) anti-P2X rabbit polyclonal antibody (Alomone) 1:200, (2) anti-P2X 2 rabbit polyclonal antibody (Abcam) 0.6 μg/mL, (3) anti-P2X 3 rabbit polyclonal antibody (Neuromics) 1:1,000, (4) anti-P2X 4 whole serum rabbit antibody (Abcam) 1:300, (5) anti-P2X 5 rabbit polyclonal antibody (Alomone) 1:200, (6) anti-P2X 6 rabbit polyclonal antibody (Abcam) 10 μg/mL or (7) anti-P2X 7 rabbit polyclonal antibody (Alomone) 1:200.

Techniques: Sequencing, Amplification

Journal: Purinergic Signalling

Article Title: Distribution of purinergic P2X receptors in the equine digit, cervical spinal cord and dorsal root ganglia

doi: 10.1007/s11302-013-9356-5

Figure Lengend Snippet: Summary of Western blot analysis plus predicted sizes for P2X 1–3, 7 proteins

Article Snippet: Membranes were blocked in 5 % skimmed dry milk in phosphate-buffered saline for 1 h at room temperature and incubated overnight at 4 °C with the primary antibody in 0.1 % Tween blocking solution, (1) anti-P2X rabbit polyclonal antibody (Alomone) 1:200, (2) anti-P2X 2 rabbit polyclonal antibody (Abcam) 0.6 μg/mL, (3) anti-P2X 3 rabbit polyclonal antibody (Neuromics) 1:1,000, (4) anti-P2X 4 whole serum rabbit antibody (Abcam) 1:300, (5) anti-P2X 5 rabbit polyclonal antibody (Alomone) 1:200, (6) anti-P2X 6 rabbit polyclonal antibody (Abcam) 10 μg/mL or (7) anti-P2X 7 rabbit polyclonal antibody (Alomone) 1:200.

Techniques: Western Blot, Molecular Weight