LPS-stimulated RAW 264.7 cells were treated with 12.5 or 25 μg/mL of P. deltoides leaf extract, after which the cells were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min. The cells were washed 3 times with PBS containing 0.5 mM MgCl2 and 1 mM CaCl2 for 5 min. After permeabilization with 0.1% Triton X-100 for 10 min, samples were incubated with blocking buffer (5% BSA in PBS). The cells were incubated with primary antibodies (p-NF-κB p65) (1:200) in 1% BSA for overnight at 4 °C. The cells were washed 3 times in PBS for 5 min and were stained for another 3 h with goat anti-rabbit IgG Texas red (1:1000) (Santa Cruz Biotechnology, Dallas, TX, USA). Cells were counterstained with 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) (Bio-Rad, Hercules, CA, USA) for 10 min to label the nuclei. The prepared cells were then observed under a confocal microscope (Olympus FV10i, Olympus, Tokyo, Japan), and images were recorded.
Goat anti rabbit igg texas red
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Goat anti-rabbit IgG Texas red is a secondary antibody conjugated with the Texas red fluorescent dye. It is designed to bind to rabbit primary antibodies, allowing for the detection and visualization of target proteins or antigens in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Ecl kit, by Thermo Fisher Scientific (4 mentions)
Ultracruz mounting medium, by Santa Cruz Biotechnology (2 mentions)
Jc 1 assay kit, by Thermo Fisher Scientific (2 mentions)
Neon transfection system, by Thermo Fisher Scientific (2 mentions)
Bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Integrin β1, by Cell Signaling Technology (1 mentions)
Integrin α5, by Cell Signaling Technology (1 mentions)
Pyap1 s127, by Cell Signaling Technology (1 mentions)
Dntp mixture, by Takara Bio (1 mentions)
Rnase inhibitor, by Promega (1 mentions)
P lats1 ser909, by Cell Signaling Technology (1 mentions)
Mitomycin c, by Merck Group (1 mentions)
M mlv reverse transcriptase, by Promega (1 mentions)
Embelin, by Merck Group (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Vdac1, by Santa Cruz Biotechnology (1 mentions)
C myc, by Merck Group (1 mentions)
Phospho gsk3β, by Cell Signaling Technology (1 mentions)
6 protocols using goat anti rabbit igg texas red
1
Immunofluorescence Analysis of NF-κB Activation
2
Cell Viability and Signaling Pathway Analysis
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RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Hyclone (Logan, VI, USA). The bicinchoninic acid (BCA) protein assay kit, and goat-anti mouse IgM Alexa Flour 488 were purchased from Thermo Scientific (Waltham, MA, USA). The D-PlusTM CCK cell viability assay kit was purchased from Dongin LS (Seoul, Korea). Mitomycin C was obtained from Sigma (St. Louis, MO, USA). M-MLV reverse transcriptase and RNase inhibitor were purchased from Promega (Madison, WI, USA). dNTP mixture was obtained from TaKaRa Bio (Shiga, Japan). QGreenTM 2X SybrGreen qPCR Master Mix was purchased from CellSafe (Gyeonggi, Korea). Rabbit poly-clonal antibodies for FAK, p-FAK (Tyr925), Src, p-Src (Tyr416), p-ERK1/2 (Thr202/Tyr204), integrin α5, integrin β1, LATS1, p-LATS1 (Ser909), and p-YAP1 (S127) were purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit polyclonal antibodies for E-cadherin, β-catenin, ERK1, and CYR61, mouse monoclonal antibodies for YAP1 and α-tubulin, and goat anti-rabbit IgG Texas-Red were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit polyclonal antibody for G0S2 was obtained from CUSABIO (College Park, MD, USA). All other chemicals and reagents were of highest quality commercially available.
3
Embelin Modulation of Apoptotic Pathways
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Embelin was purchased from Sigma-Aldrich (St. Louis, MO). A 100 mM solution of Embelin was prepared in dimethyl sulfoxide, stored as small aliquots at -20°C and then diluted as needed in cell culture medium. A 8M solution of lithium chloride was obtained from Sigma-Aldrich and diluted as needed in cell culture medium. FBS and RPMI 1640 medium were purchased from HyClone (Logan, UT). The Neon transfection system, JC-1 assay kit and COX-4 antibody were from Life Technologies (Carlsbad, CA). The bicinchoninic acid (BCA) protein assay kit and ECL kit were from Thermo Scientific (Rockford, IL). Antibodies against phospho-Akt (Ser 473), GSK-3β, phospho-GSK-3β, or Bcl-2 were from Cell Signaling Technology (Beverly, MA). Antibodies against Total-Akt, VDAC1, Bcl-xL, Bax, β-catenin, cyclin D1, GAPDH or goat anti-rabbit IgG-Texas Red and Ultra Cruz mounting medium were from Santa Cruz Biotechnology (Santa Cruz, CA). Cyclooxygenase-2 (COX-2), Mcl-1, cytochrome c, AIF, β-catenin, c-myc or histone H1 antibodies were from Millipore Co. (Bedford, MA). All other chemicals were of the highest purity or molecular biology grade and were obtained from commercial sources.
4
NFκB Activation Monitoring in P. acnes-Infected HaCaT Cells
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Heat-killed P. acnes-infected HaCaT cells were treated with Ce6-mediated PDT, after which the cells were fixed with 4% paraformaldehyde in PBS for 20 min and permeabilized with 0.5% Trixon X-100 for 15 min. After 1 h incubation with blocking buffer (5% BSA in PBS), the cells were incubated with primary antibodies (NFκB p65 Rabbit mAB) (1:100) (Cell Signaling Technology, Inc., MA, USA) in 0.5% BSA for overnight at 4°C. The cells were washed in PBS for 10 min, done 3 times, and were stained for another 1 h with goat anti-rabbit IgG Texas red (1:1000) (Santa Cruz Biotechnology, Inc., USA). Nuclei were counterstained with 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) (Bio-Rad, USA). The prepared cells were then observed under a fluorescent microscope and images were recorded.
5
Mitochondrial Apoptosis Pathway Analysis
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FBS and DMEM medium were purchased from HyClone (Logan, UT, USA). The Neon transfection system, JC-1 assay kit and cytochrome oxidase subunit (COX)-4 antibody were from Life Technologies (Carlsbad, CA, USA). The bicinchoninic acid (BCA) protein assay kit and ECL kit were from Thermo Scientific (Rockford, IL, USA). Antibodies against VDAC1, Bax, Bak, GAPDH or goat anti-rabbit IgG-Texas Red and Ultra Cruz™ mounting medium were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Cytochrome c, or AIF antibodies were from Millipore Co. (Bedford, MA, USA). All other chemicals were of the highest purity or molecular biology grade and were obtained from commercial sources.
6
LPS-Induced Inflammation in RAW 264.7 Cells
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LPS-induced inflammation in RAW 264.7 cells was examined as follows. The cells were treated with K. parviflora extract, fixed with 4% paraformaldehyde in PBS for 20 min, and permeabilized with 0.5% Triton X-100 for 15 min. After 1 h of incubation with a blocking buffer (5% BSA in PBS), the cells were incubated with primary antibodies (rabbit monoclonal antibodies against NF-κB p65; 1:100) (Cell Signaling Technology, Inc., Danvers, MA, USA) in 0.5% BSA overnight at 4 °C. The cells were washed three times with PBS for 10 min and stained for another 1 h with goat anti-rabbit IgG Texas red (1:1000) (Santa Cruz Biotechnology, Inc., Dallas, USA). Nuclei were counterstained with 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (Bio-Rad, Hercules, USA). The prepared cells were then observed under a fluorescent microscope and images were recorded [41 (link)].
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