SMAD2 (L16D3; #3103) and Snail (C15D3; #3879) antibodies were from Cell Signaling Technology. Acetylated α-tubulin (clone 6-11B-1; #T6793) antibody and Phalloidin-Atto 594 (#51927) were from Sigma-Aldrich. Antibody against α-tubulin (DM1A; #CP06) was from Calbiochem, Merck Millipore. Antibodies against p-SMAD2 and p-SMAD3 have been described previously [28 (link), 29 (link)]. Goat anti-Rabbit IgG (H + L) Alexa Fluor 488 conjugate (#A-11008), Goat anti-Mouse IgG (H + L) Alexa Fluor 488 conjugate (#A-11029), and Goat anti-Mouse IgG (H + L) Alexa Fluor 594 conjugate (A-11032) were from Life Technologies. Goat-anti-Rabbit IRDye 800CW (#926-32211) and Goat-anti-Mouse IRDye 680 (#926-32220) were from LI-COR Biosciences.
Goat anti rabbit igg h l alexa fluor 488 conjugate
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States
Goat anti-rabbit IgG (H+L) Alexa Fluor 488 conjugate is a secondary antibody labeled with the Alexa Fluor 488 fluorescent dye. It is designed to bind to rabbit primary antibodies and can be used for detection and visualization in various immunoassays and microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase, by Thermo Fisher Scientific (2 mentions)
Heme oxygenase 1, by Abcam (2 mentions)
Goat anti mouse irdye 680, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit, by LI COR (1 mentions)
Snail c15d3, by Cell Signaling Technology (1 mentions)
Phalloidin atto 594, by Merck Group (1 mentions)
Acetylated α tubulin clone 6 11b 1, by Merck Group (1 mentions)
Fluoview fv1000 clsm, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti α tubulin fitc antibody, by Merck Group (1 mentions)
Mouse monoclonal anti γ tubulin antibody, by Merck Group (1 mentions)
Cy5 conjugated goat anti human igg, by Jackson ImmunoResearch (1 mentions)
Mouse anti β actin monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Cy5 conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Fluorview 1000 confocal microscope, by Olympus (1 mentions)
Cm3050s cryostat, by Leica (1 mentions)
Oct compound, by Leica (1 mentions)
Balb c nude mice, by Shanghai Laboratory Animal Center (1 mentions)
8 protocols using goat anti rabbit igg h l alexa fluor 488 conjugate
1
Immunofluorescence Staining Protocol
2
Comprehensive Antibody Characterization for Cellular Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: Mouse anti-GAPDH, clone 6C5 #MAB374 from Sigma-Aldrich; mouse anti-TOM20 #sc-17764 (F-10), mouse anti-Lamin A/C #sc-376248 from Santa Cruz Biotechnology, Dallas, TX, USA; rabbit anti-STAT3 Y7051679131S, rabbit anti-JAK2 (Y1007/1008) 07-123, mouse anti-OPA1 #612606 from BD Transduction, San Jose CA, USA; rabbit anti-phospho DRP1 (Ser616) #3455, rabbit anti-TFAM (D5C8) #8076S from Cell Signaling Technology, Danvers, MA, USA, #8076; rabbit anti-PGC1-alpha #NBP1-04676SS from Novus; rabbit Anti-NRF1 #ab175932 from Abcam, Cambridge, UK; anti-DNA mouse monoclonal AC-30-10 #61014PROGEN from Thermo Fisher; goat anti-rabbit IgG (H + L) Alexa Fluor® 488 conjugate and goat anti-mouse IgG (H + L) Alexa Fluor® 568 conjugate from Life Technologies, Carlsbad, CA, USA.
3
Paraquat-Induced Oxidative Stress in NT2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NT2 cells were cultured on 13 mm cover slips up to 80% confluency, and then treated with 0 µM and 50 µM of paraquat. Following the treatment, the cells were fixed in 4% paraformaldehyde and permeabilized using 0.2% Triton X-100 in 1X PBS. The cells were blocked in 1% (w/v) BSA in 1X PBS and stained for overnight at 4°C with the following primary antibodies: OCT4 (1:400, #2750, Cell Signaling Technology; CST), NANOG(1:200, #3580, CST), TDGF1(1:200, #2020, CST), β3-tubulin (1:200, #4466, CST), and NeuroD1 (1:100, #4373, CST). The cells were subsequently stained with the following secondary antibodies: DyLight 488-AffiniPure Rabbit Anti-Mouse IgG (H+L) (1:200, Jackson ImmunoResearch Laboratories) and Goat Anti-Rabbit IgG (H+L) Alexa Fluor 488 Conjugate (1:200, #A11008, ThermoFisher Scientific). To measure ROS activity (superoxide level), NT2 cells were stained with 30 µM dihydroethidium (DHE) (#D1168, ThermoFisher Scientific) for 5 minutes. Slides were mounted using Vectashield mounting medium containing DAPI on a glass slide and viewed under the confocal microscope (Olympus Fluoview FV1000 cLSM).
4
Immunofluorescence and IHC Analysis of Tumor Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin-embedded tumor sections were cut at 4 µm thick and stained with H&E for pathological examination. For Ki-67 immunofluorescence, antigen retrieval was performed by heating for 20 min at 400 W in a microwave in 0.01 M citrate buffer (pH 6.0). After blocking for 45 min in 3% BSA and washing in PBS, slides were incubated overnight at 4°C with anti-Ki-67 (Thermo Scientific RM-9106-S0) and diluted 1:100 in Dako ready-to-use diluent (Dako S2022). After a PBS wash, slides were incubated with goat antirabbit IgG (H+L)-Alexa Fluor 488 conjugate (Thermo Fisher Scientific A-11008) diluted 1:200, stained with DAPI (Life Technologies D1306) diluted 1:10,000, washed once with water, and mounted with fluorescence mounting medium (Dako S3023). For anti-Omomyc immunohistochemistry, the same procedure was performed, but a primary rabbit polyclonal anti-Omomyc antibody (affinity-purified and to remove MYC cross-reactive antibodies) was used at a 0.02 mg/mL final concentration. Confocal microscopy images were captured using a Nikon C2+ confocal microscope and NIS-Elements software. Images were acquired and analyzed in a blind fashion.
5
Western Blot and Immunohistochemistry Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies (Abcam, Inc., Cambridge, MA) were used for western blotting: heme oxygenase-1 (rabbit monoclonal, clone EP1391Y), Nrf2 (rabbit monoclonal, clone EP1808Y), CD31 (rabbit monoclonal, clone EPR3094). Secondary antibody used for western blotting was polyclonal goat anti-rabbit (H+L), conjugated to horseradish peroxidase (Life Technologies, cat. # G-21234). The following antibody was used for immunohistochemical staining of Nrf2 in cells [66 (link)]: Nrf2 (H-300), sc-13032 rabbit polyclonal raised against amino acids 37–336 of human Nrf2 (Santa Cruz Biotechnology Inc.). Secondary antibody used for immunohistochemical staining was goat anti-rabbit IgG (H+L) Alexa Fluor 488 conjugate (ThermoFisher).
6
Antibody Usage for Cellular Analysis
7
Kif2a and Centromere Protein Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit polyclonal anti-Kif2a antibody used in Western blot and immunofluorescence was purchased from Novus Biologicals (Littleton,CO); Mouse monoclonal anti-α-tubulin-FITC antibody and mouse monoclonal anti-γ-tubulin antibody were obtained from Sigma-Aldrich Co. (Cat# F2168, Ca#T6557); Rabbit polyclonal anti-bub3 antibody was obtained from Santa Cruz Biotechology (Ca#sc-28258); Human polyclonal anti-centromere antibody (ACA) was obtained from Antibodies Incorporated (Item # 15-234-0001). Alexa Fluor@ 488-conjugate Goat anti-Rabbit IgG (H + L) and Alexa Fluor @594-conjugate Goat anti-Rabbit IgG (H + L) were produced by Thermo Fisher Scientific (Catalog# A-11008, Catalog# A-11012); Cy5-conjugated goat anti-human IgG and Cy5-conjugated goat anti-mouse IgG were purchased from Jackson ImmunoResearch Laboratory (West Grove, PA) (Cat#115-175-146, Cat#115-175-062). Mouse monoclonal anti-β-actin antibody was purchased from Santa Cruz Biotechnology (SC-8432, Santa Cruz, CA).
All other reagents were purchased from Sigma Aldrich except when mentioned otherwise.
All other reagents were purchased from Sigma Aldrich except when mentioned otherwise.
8
Quantifying Tumor-Associated Macrophages in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the infiltration of TAMs in vivo, 4-week-old male BALB/c nude mice were obtained from the Shanghai Laboratory Animal Center and used in our experiments. A total of 5 × 106 MDA-MB-231 cells (shcon or shCCL2 cells) were inoculated into the right flank of mice subcutaneously. EPZ-6438 was given by oral gavage 200 mg/kg/day in the treatment group as indicated. Nearly four weeks later, tumor tissues harvested from mice were frozen in Tissue-Tek O.C.T. compound and sectioned in Leica CM3050 S Cryostats, fixed in acetone, washed and blocked with PBS containing 5% goat serum. The immunodetection of F4/80 and CD206 were carried out by incubating cells with rabbit anti-CD206 and mouse anti-F4/80 primary antibody diluted 1:200 in PBS containing 3% BSA. After incubation overnight, the slides were washed three times with PBS and incubated for 1 h at room temperature with Alexa Fluor 488-conjugate goat anti-rabbit IgG (H + L) and Alexa Fluor 633-conjugate goat anti-mouse IgG (H + L) secondary antibody (ThermoFisher Scientific, USA) diluted 1:200 in PBS containing 3% BSA. Nuclei were stained using 4ʹ,6-diamidino-2-phenylindole (DAPI). The fluorescence signals were analyzed using an Olympus Fluorview 1000 confocal microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!