The immunofluorescence assay was performed as previously described (13 (link), 29 (link)). Human lung sections, rats’ lung sections (13 (link)), and HPASMCs were first stained with anti-α-smooth muscle actin (αSMA, Boster, Wuhan), anti-MBD2 (Abcam), and anti-Ki67 primary antibodies (Abcam), followed by staining with Alexa Fluor 594-labeled anti-mouse/rabbit, or Alexa Fluor 488-conjugated anti-rabbit/mouse antibodies (Invitrogen, Carlsbad, CA, USA), respectively. DAPI was used for staining the cell nuclei. A Pannoramic MIDI (3Dhistech, Budapest, Hungary) was used to scan and analyze the images. Mean fluorescence intensity was assessed by ImageJ software (NIH, Bethesda, MD).
Alexa fluor 594 labeled anti mouse rabbit
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 594-labeled anti-mouse/rabbit is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 594. It is designed to detect and visualize primary antibodies raised in mouse or rabbit.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using alexa fluor 594 labeled anti mouse rabbit
1
Immunofluorescence Assay for Lung Tissue Analysis
2
Histological Analysis of Lung Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human lung tissue and the left lung of mice were inflated in fresh 4% paraformaldehyde for 24 h at room temperature. Then, the lung tissue was embedded in paraffin and sliced into 5 μm sections. The sections were subjected to hematoxylin and eosin (H&E), Sirius red and Masson's trichrome staining. The Ashcroft scores of the mice were determined to assess the severity of lung fibrosis in each mouse by a blinded observer according to the established protocol.18 For immunofluorescence staining, BALF cytospin slides or paraffin sections were probed with antibodies against CD68, MECP2, and IRF4, followed by staining with Alexa Fluor 594–labeled anti‐mouse/rabbit or Alexa Fluor 488–conjugated anti‐rabbit/mouse antibodies (Invitrogen).
3
Histological Analysis of Interstitial Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The left lung was inflated and placed in fresh 4% neutral-buffered paraformaldehyde for 24 hours at room temperature, embedded in paraffin, and subjected to the histological analysis as previously reported (17 (link)). Each successive field was individually assessed for the severity of interstitial fibrosis in a blinded fashion by two pathologists using the Ashcroft scoring system (46 (link)). For immunostaining, the frozen sections (7 μm) were probed with antibodies against CD206, CD68, Mbd2, and F4/80, followed by staining with Alexa Fluor 594–labeled anti-mouse/rabbit or Alexa Fluor 488–conjugated anti-rabbit/mouse antibodies (Invitrogen, Carlsbad, CA, USA), respectively. COVID-19 viral immunostaining was conducted as previously reported (47 (link)).
4
Lung Tissue Histological Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The left lung was inflated in fresh 4% neutral-buffered paraformaldehyde for 24 hours at room temperature. The lung tissue was next embedded in paraffin and sliced into 5-μm sections. The sections were subjected to hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) staining using the established techniques (7 (link)). Each successive field was individually assessed for the severity of peribronchial inflammation and analyzed for mucus-containing cells by two pathologists in a blinded fashion. For immunofluorescence staining, BALF cytospin slides or sections were probed with antibodies against CD14, CD206, Arg-1, Mbd2, and F4/80, followed by staining with Alexa Fluor 594-labeled anti-mouse/rabbit or Alexa Fluor 488–conjugated anti-rabbit/mouse antibodies (Invitrogen, Carlsbad, CA, USA). For immunohistochemistry (IHC) staining of Arg-1, the IHC staining kit was purchased from Servicebio (Wuhan, China).
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!