After blocked with BSA for 30 minutes, sections (5 μm) of abdominal arteries were incubated with rabbit anti‐COX‐2 (1:100, Abcam), mouse anti‐CD68 (1:100, Abcam) or mouse anti‐α‐SM actin (1:100, Sigma‐Aldrich) overnight at 4°C. After incubated with secondary antibodies, a drop of Prolong Gold anti‐fade reagent with DAPI (Vector Laboratories) was used to seal the coverslip. Images were acquired by laser scanning confocal microscopy (LSM 710, Zeiss).
Prolong gold anti fade reagent with dapi
Manufactured by Vector Laboratories
Sourced in United States
Prolong Gold anti‐fade reagent with DAPI is a mounting medium designed to retard the photobleaching of fluorescently labeled specimens. It contains the nuclear counterstain DAPI, which binds to DNA and emits blue fluorescence.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710, by Zeiss (3 mentions)
Mouse anti cd68, by Abcam (1 mentions)
Rabbit anti cox 2, by Abcam (1 mentions)
Mouse anti α sm actin, by Merck Group (1 mentions)
Mouse monoclonal anti icam 1, by Santa Cruz Biotechnology (1 mentions)
Rabbit monoclonal anti hmgb1, by Abcam (1 mentions)
Alexa 594 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
μ slide, by Ibidi (1 mentions)
Lab tek chamber slide, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions)
Perm wash buffer, by BD (1 mentions)
Superfrost plus slide, by Thermo Fisher Scientific (1 mentions)
Saponin, by BD (1 mentions)
Fv100mpe, by Olympus (1 mentions)
7 protocols using prolong gold anti fade reagent with dapi
1
Immunohistochemical Analysis of Abdominal Arteries
2
Immunocytochemistry of Cell Transfection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated into Millicell EZ SLIDE 8‐well glass (Millipore PEZGS0816) at a density of 2 ×104 cells/well and transfected with gapmers (Table S1 ) at 50 nM for 24 h. Cells were fixed in 4% PFA and blocked with blocking buffer (PBS/5% normal serum/0.3% Triton™ X‐100) at room temperature for 60 min. Cells on slides were incubated with primary antibody at the dilutions indicated in the next section in antibody dilution buffer (PBS/1% BSA/0.3% Triton™ X‐100) at 4°C overnight. After primary antibody incubation, the cells were incubated with fluorochrome‐conjugated secondary antibodies according to the manufacturer's instructions at room temperature for 1 h in the dark. The slides were then washed with PBS three times for 10 min each, then mounted with Prolong® Gold Antifade Reagent with DAPI (Vector Laboratories, H‐1200‐10).
3
Immunofluorescence Imaging of HMGB1 and ICAM-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After stimulation, HUVECs were fixed in 4% paraformaldehyde and permeabilized in 0.1% Triton X-100/PBS for 5 min. After being blocked with BSA for 30 min, cells were incubated with the primary antibodies rabbit monoclonal anti-HMGB1 (1:1000, Abcam) and mouse monoclonal anti-ICAM-1 (1:500; Santa Cruz Biotechnology) overnight at 4°C. After being washed with TBS-T, cells were incubated with secondary antibodies (1:500; Jackson immunoresearch). A drop of Prolong Gold antifade reagent with DAPI (Vector Laboratories, CA, USA) was used to seal the coverslip. Images were acquired by laser scanning confocal microscopy (LSM 710; Carl Zeiss, Germany). Data were analyzed by use of Image-Pro Plus 6.0.
4
Quantifying NF-κB Activation in Vascular Smooth Muscle
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vascular smooth muscle cells were cultured, transfected with si‐NONO and stimulated with Ang II as explained earlier. Cells were fixed using 4% paraformaldehyde for 20 minutes and permeabilized in PBS with 0.1% Triton X‐100. Following the blocking with goat serum for 30 minutes at RT, samples were incubated with rabbit anti‐NF‐κB p65 (Cell Signaling Technology, 8424, 1:100) and mouse anti‐NONO (Santa Cruz, sc‐166702x, 1:100) antibodies overnight at 4°C. Alexa 488‐conjugated goat anti‐mouse IgG and Alexa 594‐conjugated goat anti‐rabbit IgG (Invitrogen) were used as the secondary antibodies. IgG and secondary antibodies were considered as negative controls. A droplet of Prolong Gold anti‐fade reagent with DAPI (Vector Laboratories) was added to fix the coverslip. Images were captured via laser scanning confocal microscopy (LSM 710, Zeiss).
5
Immunofluorescence Staining of Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U2OS, HeLa, and NIH3T3 cells were plated on LabTek™ chamber slides (Thermo Fisher Scientific), and mESCs were plated on μ-Slide (ibidi). Cells were pre-extracted with CSK buffer (10 mM PIPES, 100 mM NaCl, 300 mM sucrose, 3 mM MgCl2, 1 mM EGTA, and 0.5% Triton X-100) for 10 min at 4°C and fixed with 4% paraformaldehyde for 20 min at room temperature. The fixed cells were incubated with the primary antibodies diluted in PBS supplemented with 10% fetal bovine serum for 1 h at room temperature. After three washes with 0.05% Triton X-100 in PBS, Alexa Fluor®–conjugated secondary antibodies (Thermo Fisher Scientific) were added and incubated for 30 min. Cells were mounted using ProLong® Gold antifade reagent with DAPI (Vector Laboratories).
6
Germ Cell Identification in Chick Testis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed to identify the expression of GFP and DAZL in germ cells of DAZL::GFP chick testis. Testes of chicks at hatching day were paraffin‐embedded and sectioned (thickness, 9–10 μm). After deparaffinization, sections were washed three times with PBS and blocked with a blocking buffer (5% goat serum and 1% bovine serum albumin in PBS) for 1 h at room temperature. Sections were then incubated at 4 °C overnight with primary antibodies, rabbit anti‐GFP (Invitrogen, Thermo Fisher Scientific Inc., Carlsbad, CA, USA), or rabbit anti‐DAZL [16 (link)]. After washing three times with PBS, sections were incubated with fluorescence‐conjugated secondary antibodies (Alexa Fluor 488 or 568) for 1 h at room temperature. After washing three times, sections were mounted with ProLong Gold antifade reagent with DAPI (Vector Laboratories, Burlingame, CA, USA) and visualized on a fluorescence microscope.
7
Immunofluorescence analysis of recombinant CL-K1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CHO DG44 cells expressing and secreting recombinant full-length CL-K1 (see above) were cultivated in suspension in PowerCHO 2 serum free media (Lonza). The cells were transferred by cytospin onto Superfrost plus slides (Thermo Scientific) coated with polylysine. Cells were fixed with 1% PFA in PBS for 10 min at RT. After fixation, the cells were washed thrice with PBS before permeabilization with BD Perm/wash buffer containing Saponin (Becton Dickinson) for 15 min at RT. After permeabilization, cells were rinsed thrice with PBS and blocked for 1 hr. with PBS, 1% BSA and 0.3 % Trition-X-100. After blocking, cells were incubated with 1 μg Alexa 488 conjugated mAb 16-25 per ml blocking buffer and incubated for 2h at RT. The mAb:Alexa ratio was 1:25 and the conjugation was performed with (non-biotinylated) mAb in solution and not on beads. After incubation, the cells were rinsed in the blocking buffer, coverslips were mounted with Prolong Gold Antifade Reagent with DAPI (Vector Laboratories) and dried ON, before imaging on a confocal laser-scanning microscope (Olympus FV100 MPE). A 488 nm laser line was used to excite Alexa 488 mAb and 405 nm for DAPI.
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!