Immunofluorescence Staining of Cleaved Caspase 3

Immunofluorescence was performed as recently described^{29 (link)}. Cells were fixed with 3% formaldehyde/PBS for 30 min, permeabilized with 0.5% Triton X-100/PBS for 10 min and blocked with 3% BSA/PBS for 1 h. Staining of cleaved caspase 3 (Cell Signaling, #9664) was carried out overnight at 4 °C and with anti-rabbit secondary antibody for 2 h at room temperature. After several washes with PBS, samples were covered with Vectashield/DAPI mounting medium (Vector Labs). Images were acquired using an AxioImager.Z1/ApoTome microscope (Zeiss).

Free full text: Click here

Eke I., Aryankalayil M.J., Bylicky M.A., Makinde A.Y., Liotta L., Calvert V., Petricoin E.F., Graves E.E, & Coleman C.N. (2022). Radiotherapy alters expression of molecular targets in prostate cancer in a fractionation- and time-dependent manner. Scientific Reports, 12, 3500.

Publication 2022

cleaved caspase 3 Vectashield/DAPI mounting medium AxioImager.Z1/ApoTome microscope

Anti antibody Caspase 3 Cells Dapi Formaldehyde Immunofluorescence Microscope Rabbit Triton x 100 Vector

Corresponding Organization :

Other organizations : Stanford University, National Cancer Institute, National Institutes of Health, Center for Cancer Research, George Mason University

Top 5 similar protocols

Variable analysis

independent variables

Immunofluorescence protocol

dependent variables

Cleaved caspase 3 staining

control variables

Formaldehyde/PBS fixation
Triton X-100/PBS permeabilization
BSA/PBS blocking
Anti-rabbit secondary antibody staining
Vectashield/DAPI mounting medium

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!