Bone slides were also processed for immunohistochemistry (IHC). IHC slides were deparaffinized, hydrated and endogenous peroxidase blockade was performed with 0.3 % hydrogen peroxide. Antigen retrieval was performed with 0.1 M citrate buffer solution. The sections were then coated with primary rabbit anti-mouse cleaved Caspase-3 antibody (1:300; Cell Signaling Technology (Asp175 - #9961)), for labeling cleaved Caspase-3. Then the sections were incubated with the secondary antibody (VECTASTAIN Elite ABC HRP Kit) and stained with DAB chromogenic solution (Sigma-Aldrich). Counterstaining with hematoxylin was performed. Negative reaction controls comprised samples in which the primary antibodies were omitted.
Anti mouse cleaved caspase 3 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-mouse cleaved Caspase-3 antibody is a laboratory reagent used to detect the presence of the cleaved form of Caspase-3 protein in mouse samples. Caspase-3 is a key enzyme involved in the apoptosis (programmed cell death) pathway. The antibody specifically recognizes the cleaved, activated form of Caspase-3.
Automatically generated - may contain errors
Lab products found in correlation
Vectastain elite abc hrp kit, by Merck Group (1 mentions)
Anti mouse cd31, by BD (1 mentions)
Goat anti rat cy3, by Jackson ImmunoResearch (1 mentions)
Anti mouse pd l1 antibody, by BioXCell (1 mentions)
Alexa fluor 488 goat anti rabbit, by Jackson ImmunoResearch (1 mentions)
Anti mouse cd31 antibody, by BD (1 mentions)
3 3 diaminobenzidine kit, by Vector Laboratories (1 mentions)
Goat anti rat secondary antibody, by Jackson ImmunoResearch (1 mentions)
Rabbit anti mouse tnf α antibody, by Abcam (1 mentions)
Mouse anti gfap antibody, by Merck Group (1 mentions)
Confocal microscope, by Leica (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Tunel apoptosis detection kit, by Roche (1 mentions)
Fluoroshield with dapi, by Merck Group (1 mentions)
Iba1 antibody, by Fujifilm (1 mentions)
Mouse anti nlrp3 antibody, by Thermo Fisher Scientific (1 mentions)
Rabbit anti neun antibody, by Merck Group (1 mentions)
Rabbit anti collagen 1 antibody, by Abcam (1 mentions)
Rabbit anti c3 antibody, by Abcam (1 mentions)
5 protocols using anti mouse cleaved caspase 3 antibody
1
Immunohistochemical Analysis of Cleaved Caspase-3
2
Tissue-Specific Immunohistochemistry Analysis
3
Evaluating Treatment-Induced Toxicity and Tumor Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate potential toxicity of the treatment, B16-F10 tumor and mouse organs
(heart, liver, spleen, lung and kidney) were harvested and fixed with buffered
4% paraformaldehyde for 72 h and embedded in paraffin wax.
4 μm sections were stained with hematoxylin and eosin (H&E).
Histological sections of E.G7-OVA tumors in the therapeutic vaccine experiment
were also stained with H&E.
For immunohistochemistry, tumor sections were deparaffinized, rehydrated in
graded series of alcohol and antigen retrieval was performed in citrate buffer
(pH 6.0) for 3 minutes in an autoclave. Endogenous peroxidase activity
was inhibited by incubation with 3% H2O2 for
15 min in the dark. After blocking nonspecific reactivity with normal
goat serum for 40 min at 37 °C, samples were incubated
overnight at 4 °C with rabbit anti-mouse TNF-α antibody
(Abcam, 1:100) or anti-mouse cleaved caspase-3 antibody (Cell signaling
technology, 1:300), followed by the incubation with biotinylated goat
anti-rabbit secondary antibody at 37 °C for 40 min and
streptavidin-biotin complex at 37 °C for another 40 min.
The immunoreaction was developed using diaminobenzidine peroxide solution. Cell
nuclei were gently counterstained with hematoxylin. Images were obtained with a
Leica DM 2500 microscope.
(heart, liver, spleen, lung and kidney) were harvested and fixed with buffered
4% paraformaldehyde for 72 h and embedded in paraffin wax.
4 μm sections were stained with hematoxylin and eosin (H&E).
Histological sections of E.G7-OVA tumors in the therapeutic vaccine experiment
were also stained with H&E.
For immunohistochemistry, tumor sections were deparaffinized, rehydrated in
graded series of alcohol and antigen retrieval was performed in citrate buffer
(pH 6.0) for 3 minutes in an autoclave. Endogenous peroxidase activity
was inhibited by incubation with 3% H2O2 for
15 min in the dark. After blocking nonspecific reactivity with normal
goat serum for 40 min at 37 °C, samples were incubated
overnight at 4 °C with rabbit anti-mouse TNF-α antibody
(Abcam, 1:100) or anti-mouse cleaved caspase-3 antibody (Cell signaling
technology, 1:300), followed by the incubation with biotinylated goat
anti-rabbit secondary antibody at 37 °C for 40 min and
streptavidin-biotin complex at 37 °C for another 40 min.
The immunoreaction was developed using diaminobenzidine peroxide solution. Cell
nuclei were gently counterstained with hematoxylin. Images were obtained with a
Leica DM 2500 microscope.
4
Microglial Cytokine Effects on Neural Stem Cell Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neural stem/progenitor cells (NSPCs) were generated from the subependymal ventricular zone (SVZ) of young adult mice and cultured under clonal conditions, according to previously published procedures (Zhang et al., 2021 (link)). Microglia were plated at a density of 5 × 105 cells/cm2 and treated with each of the cytokines as described in Section “Stimulation of Primary Microglia,” followed by washing with PBS twice, and then addition of DMEM-F12 + GlutaMax medium for another 24 h. The microglial medium was collected and used as conditioned medium to culture NSPCs for 24 h. Apoptosis of NSPCs were labeled overnight using the mouse anti-cleaved caspase-3 antibody (1:300; Cell Signaling Technology, United States), followed by the anti-mouse secondary antibodies mentioned in Section “Immunofluorescence.” Finally, cells were stained for 5 min with DAPI (1:10,000) and imaged using a fluorescence microscope (Olympus BX51, Japan).
5
Multimodal Neuroinflammation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sections were boiled in citric acid buffer and were treated with 0.3% (v/v) Triton X-100 and 10% (v/v) goat serum, they were incubated overnight at 4 °C with primary antibodies (rabbit anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody, Wako, Japan; mouse anti-NLRP3 antibody, Thermo Fisher, USA; mouse purified anti-β-amyloid, 1-42 antibody, BioLegend, USA; mouse purified anti-β-amyloid, 1-40 antibody, BioLegend, USA; rabbit anti-collagen I antibody, Abcam, USA; rat anti-LAMP2 antibody, Abcam, USA; mouse anti-GFAP antibody, Sigma, USA; rabbit anti-C3 antibody, Abcam, USA; rabbit anti-NeuN antibody, Millipore, USA; mouse anti-cleaved caspase-3 antibody, Cell Signaling Technology, USA) and then incubated with secondary antibodies. Apoptosis was detected using a transferase-mediated deoxyuridine triphosphate-biotin nick end labeling Kit (TUNEL Apoptosis Detection Kit, Roche, Switzerland). Slices were embedded using Fluoroshield™ with DAPI (Sigma, USA). Images were acquired using a Nikon fluorescence microscope (Nikon, Japan) or a confocal microscope (Leica, Germany).
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!