Cells were fixed with 4% paraformaldehyde for 20 min, washed with PBS, blocked with 1% BSA (bovine serum albumin) in PBS (blocking solution) for 30 min, and permeabilized with 0.1% Triton X100 for 3 min. Primary antibodies were diluted in blocking solution and incubated with cells at room temperature for 1 h. After three washes with PBS, cells were incubated for 1 h with secondary antibodies (diluted in PBS to final concentration 4 μg/ml). After three washes with PBS, cell nuclei were stained with Hoechst. The primary antibodies used for immunocytochemistry were rat anti-MBP (Serotec) used to measure OPC differentiation, and rabbit anti-AcH3K14 (against acetylated lysine 14 in histone H3, Millipore). Secondary antibodies were goat anti-rabbit IgG Alexa Fluor 488, and rabbit anti-rat IgG Alexa Fluor 488 (Invitrogen).
Rabbit anti rat igg alexa fluor 488
Manufactured by Thermo Fisher Scientific
Sourced in United States
Rabbit anti-rat IgG Alexa Fluor 488 is a secondary antibody conjugated with the Alexa Fluor 488 fluorescent dye. It is designed for the detection and quantification of rat immunoglobulin G (IgG) in various applications, such as immunoassays and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti mbp, by Bio-Rad (3 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 546 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Tcs sp2, by Leica (1 mentions)
Antifade solution, by Vector Laboratories (1 mentions)
Alexa fluor 647 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Fitc goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Tcs sp5 mp, by Leica (1 mentions)
5 protocols using rabbit anti rat igg alexa fluor 488
1
Immunocytochemical Analysis of OPC Differentiation
2
Immunofluorescence Staining of MBP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with 4% paraformaldehyde, washed with PBS, permeabilized with 0.1% Triton X-100 for 5 min, and blocked with 1% bovine serum albumin in PBS and 0.1% Triton-100 (blocking solution) for 1 h. Primary antibodies (rat anti-MBP, 1:200 dilution, Serotec) were diluted in blocking solution and incubated at room temperature for 1 h. Samples were washed 3 times with PBS and incubated with secondary antibodies (rabbit anti-rat IgG Alexa Fluor 488, 1:200 dilutions, Invitrogen) in PBS for 1 h, followed by washing and staining of nuclei with Hoechst 33342 at a 1:1000 dilution for 5 min.
3
Immunofluorescent Staining of Oligodendrocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 7
Cells were fixed with 4% paraformaldehyde, washed with PBS, permeabilized with 0.1% Triton X-100 for 5 min, and blocked with 1% bovine serum albumin in PBS and 0.1% Triton-100 (blocking solution) for 1 h. Primary antibodies (rat anti-MBP, 1:200 dilution, Serotec) were diluted in blocking solution and incubated at room temperature for 1 h. Samples were washed 3 times with PBS and incubated with secondary antibodies (rabbit anti-rat IgG Alexa Fluor 488, 1:200 dilutions Invitrogen) in PBS for 1 h, followed by washing and staining of nuclei with Hoechst 33342 at a 1:1000 dilution for 5 min.
4
Immunofluorescence Staining of Foxp3 and IL-17
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To perform immunofluorescence staining, the sections were firstly deparaffinized in xylenes using three changes for 5 min each and then hydrated gradually through graded alcohols: wash in 100% ethanol twice for 10 min each and then 95% ethanol twice for 10 min each. Antigen retrieval was performed by heating the slides in 10 mM sodium citrate buffer, pH 6.0, at 95°C for 5 min and then cooling them in the buffer for approximately 20 min until they reached room temperature. Afterward, the slices were washed with PBS three times and the excess liquid was aspirated. After several blocking steps, the sections were incubated with 1:100 rabbit anti-mouse Foxp3 mAb (H-190, Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and 1:100 rat anti-mouse IL-17 mAb (TC11-18H10, Santa Cruz) at 37°C for 1 hr and then at 4°C overnight. The slices were washed with PBS three times for 5 min each and then incubated with Alexa Fluor 546 donkey anti-rabbit IgG (Invitrogen, CA, USA) and Alexa Fluor 488 rabbit anti-rat IgG (Invitrogen) for 2 hrs. Finally, the sections were mounted with glycerol and examined under laser scanning confocal microscopy (TCS SP2, Leica, Germany). The quantitative analysis of Foxp3+ cells and IL-17+ cells was also performed as mentioned above.
5
Immunofluorescent Staining of Circulating Tumor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CTCs slides were fixed with ethyl alcohol for 10 min at room temperature and incubated overnight at 4°C with mouse anti-human CK (to detect normal and neoplastic pancreatic cells) monoclonal antibodies (dilution at 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, United States) along with rat anti-human CD45 (to detect hematologic cells; dilution at 1:50; Santa Cruz Biotechnology, Inc.) or rabbit anti-human KLF8 (dilution at 1:100; Abcam, Cambridge, United Kingdom) and rat anti-human vimentin (dilution at 1:100; Santa Cruz Biotechnology, Inc.). The following fluorescence-conjugated secondary antibodies were used: Cy5 rabbit anti-mouse IgG (dilution at 1:400; Pierce of Thermo Fisher Scientific), and Alexa Fluor 488 rabbit anti-rat IgG, Alexa Fluor 647 goat anti-mouse IgG, Cy3 goat anti-rat IgG, or FITC goat anti-rabbit IgG (dilution at 1:400; Invitrogen). All slides were subsequently stained with DAPI (dilution at 1:1000; Pierce) and coverslipped in antifade solution (Vector Laboratories, Inc., Burlingame, CA, United States). Slides containing a mixture of human PBMCs and AsPC-1 cells were used for positive controls. The slides were imaged on a confocal laser-scanning microscope (Leica TCS SP5 MP; Leica Microsystems, Wetzlar, Germany). Cell counts are expressed as the number of cells per 7 mL of blood.
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!