Spinal cord tissue segments (S3, injury section) from the injury site were fixed with 4% paraformaldehyde (PFA) in phosphate-buffered saline (PBS) overnight(Haque et al., 2017a (link)). Tissue was then embedded in paraffin, sectioned, and mounted onto slides for analysis. Following epitope retrieval, non-specific binding sites were blocked with the serum of the secondary antibody host for 1h at room temperature (Samantaray et al., 2015 (link)). Tissues were incubated with vimentin (1:500, Abcam ab92547), Iba1 (1:250, Abcam ab5076), GFAP (1:250, Santa Cruz sc-9065), Cox-2 (1:100, Abcam ab15191), caspase-1 (1:200, Abcam ab74279), CSPG (1:200, Sigma C8035), or NFP (1:1000, Sigma Aldrich N4142) antibodies overnight at room temperature. Sections were then incubated with Texas Red® Goat Anti-Rabbit IgG Antibody (Vector Laboratories, Inc., Burlingame, CA), DyLight® 488 Anti-Goat IgG (H+L) (Vector Laboratories, Inc., Burlingame, CA), Texas Red® Anti-Mouse IgM Antibody (Vector Laboratories, Inc., Burlingame, CA), or Fluorescein Anti-Rabbit IgG (Vector Laboratories, Inc., Burlingame, CA) for 1h in the dark. Slides were mounted with 1 drop of Invitrogen™ ProLong™ Gold Antifade Mountant with DAPI (Thermo Fisher Scientific) and coverslipped. After staining, SCI tissues were viewed under a fluorescence microscope with representative images taken at 20× magnification.
Fluorescein anti rabbit igg
Manufactured by Vector Laboratories
Fluorescein Anti-Rabbit IgG is a secondary antibody conjugated with the fluorescent dye fluorescein. It is designed to detect and visualize rabbit immunoglobulin G (IgG) in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Invitrogen prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Fluoview fv1000 confocal microscope, by Olympus (1 mentions)
Normal horse serum, by Vector Laboratories (1 mentions)
Normal goat serum (ngs), by Vector Laboratories (1 mentions)
Normal horse serum, by Merck Group (1 mentions)
Gfap cy3 antibody, by Merck Group (1 mentions)
Confocal microscope, by Zeiss (1 mentions)
Mitotracker deep red 633, by Thermo Fisher Scientific (1 mentions)
3 protocols using fluorescein anti rabbit igg
1
Immunohistochemical Analysis of Spinal Cord Injury
2
Immunostaining for Brain Tumor Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunostaining was performed using the previously established protocol in our lab [19 (link)]. Frozen 25 μm coronal sections encompassing the entire tumor were generated from each mouse brain. The sections were blocked with 5% normal goat serum/5% normal horse serum (Vector lab., Burlingame, CA) in PBS containing 0.3% Triton X-100 and 0.05% Phenylhydrazine for 30 minutes and then incubated with monoclonal mouse anti-Pyk2 antibody, dilution 1:1000 (Cell Signaling; #3480S), polyclonal rabbit anti-Iba1 antibody (Wako; #019–19741) 1 μg/ml, and monoclonal GFAP-Cy3 antibody produced in mouse (Sigma, #C9205), dilution 1:500, in PBS-TAT (0.3% TritonX-100, 5% normal goat/5% normal horse serum, 1% sodium azide, 0.01% thimerosal) overnight at 4°C. The sections were incubated with corresponding secondary antibodies (AMCA anti-mouse IgG and fluorescein anti-rabbit IgG (Vector Lab., Burlingame, CA)) overnight and visualized using an Olympus Fluoview FV1000 confocal microscope with 10× objectives or 40× oil immersion objectives.
3
Immunofluorescent Localization of p38β
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After fixation and permeabilization, NRCM (1×104 cells/mL) or female heart tissue sections (in 5 μm thickness) were stained with anti-p38β primary antibody (sc-6187-R, Santa Cruz) at room temperature for 1 hr. Following triple wash in PBS, they were incubated with Fluorescein anti-rabbit IgG (V0729, Vector laboratories) for 30 min in dark, rinsed in PBS three times, then co-incubated with 1 mM MitoTracker deep red 633 (Molecular Probes) to stain mitochondria. Heart tissue slices were mounted with mounting medium containing 4’,6-diamidino-2-phenylindole (DAPI), and observed under confocal microscope (Zeiss) for p38β (green fluorescence), mitochondria (red) and nucleus (blue).
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!