The hippocampus was removed and homogenized in RIPA buffer with protease inhibitor cocktail (Nacalai tesque). The homogenate was centrifuged at 15,000 rpm for 10 min at 4 °C. The supernatants were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (e-PAGEL-HR, ATTO), and proteins transferred onto an Immobilon-P membrane (Clear Blot Membrane-P plus, ATTO). The membranes were blotted with antibodies of anti-Iba-1 (019–19,741, rabbit, 1:500, Wako), anti-synaptophysin (ab32127, rabbit, 1:1000, Abcam), anti-PSD95 (ab18258, rabbit, 1:10,000, Abcam) and anti-β-actin (G043, mouse, 1:1000, Abm) antibodies. The sections were treated with secondary antibody (NA931 and NA934, 1:10,000, GE Healthcare) and protein bands visualized with a chemiluminescence detection system (ECL Select Western Blotting Detection Reagent, Amersham). Immunoblot signals were analyzed by a LAS-4000 digital imaging system (Fujifilm, Tokyo, Japan). Band intensities were quantified and divided by their corresponding loading controls (β-actin) (n = 3 animals per group).
019 19 741 rabbit
Manufactured by Fujifilm
Sourced in Japan
019–19,741, rabbit is a laboratory equipment product manufactured by Fujifilm. It is designed to perform specific laboratory functions, but a detailed description cannot be provided while maintaining an unbiased and factual approach. More information may be available from the manufacturer.
Automatically generated - may contain errors
Lab products found in correlation
Ab18258, by Abcam (1 mentions)
Ab32127, by Abcam (1 mentions)
Protease inhibitor cocktail, by Nacalai Tesque (1 mentions)
Na931, by GE Healthcare (1 mentions)
Na934, by GE Healthcare (1 mentions)
Ecl select western blotting detection reagent, by Cytiva (1 mentions)
Las 4000 digital imaging system, by Fujifilm (1 mentions)
Ab150079, by Abcam (1 mentions)
Fv3000, by Olympus (1 mentions)
2 protocols using 019 19 741 rabbit
1
Hippocampal Protein Expression Analysis
2
Quantification of Astrocyte and Microglia Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescence staining, sections from wild-type, littermates and CAMKII-Cre:Cbsfl/fl mice were washed and incubated in 5% donkey serum or 5% goat serum
separately in 1x PBS to block non-specific sites. After 2 h, sections were incubated in the GFAP antibody (1:400, ab4674-chicken, Abcam) and Iba1 antibody (1:400, 019-19741-rabbit, Wako,
Osaka, Japan) solution, respectively, at 4°C for overnight. After three rinses, sections were incubated in Alexa Fluor 647 conjugated donkey anti-chicken IgY (1:200, 703-165-155, Jackson
Immunoresearch, West Grove, PA, USA) and Alexa Fluor 647 conjugated goat anti-rabbit IgG (1:200, ab150079, Abcam) solution, respectively, for 2 h at room temperature. Finally, sections were
washed and mounted onto slides, followed by coverslipping with a fluorescent mounting medium. Fluorescent sections were imaged with an Olympus confocal microscope (FV3000, Olympus, Tokyo,
Japan). For each mouse, three random fields were selected from each section. The fluorescence intensity of astrocytes and microglial cells was quantified using the ImageJ software (NIH).
separately in 1x PBS to block non-specific sites. After 2 h, sections were incubated in the GFAP antibody (1:400, ab4674-chicken, Abcam) and Iba1 antibody (1:400, 019-19741-rabbit, Wako,
Osaka, Japan) solution, respectively, at 4°C for overnight. After three rinses, sections were incubated in Alexa Fluor 647 conjugated donkey anti-chicken IgY (1:200, 703-165-155, Jackson
Immunoresearch, West Grove, PA, USA) and Alexa Fluor 647 conjugated goat anti-rabbit IgG (1:200, ab150079, Abcam) solution, respectively, for 2 h at room temperature. Finally, sections were
washed and mounted onto slides, followed by coverslipping with a fluorescent mounting medium. Fluorescent sections were imaged with an Olympus confocal microscope (FV3000, Olympus, Tokyo,
Japan). For each mouse, three random fields were selected from each section. The fluorescence intensity of astrocytes and microglial cells was quantified using the ImageJ software (NIH).
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!