Apoptosis post exposure was investigated by western blot analysis of cleaved and uncleaved caspase‐3. Cells were lysed in 200 μl 1× Laemmli buffer and boiled at 95°C for 10 min. Samples were stored at −20°C until analysis was performed. Cell lysates and caspase‐3 control extracts (#9663; Cell Signalling Technology) were loaded onto 4%–20% gradient polyacrylamide gels (Bio‐Rad). Proteins were separated by gel electrophoresis, transferred to PVDF membranes (Bio‐Rad) and incubated with appropriate antibodies. Primary antibodies were caspase‐3 antibody (#9662; Cell Signalling Technology) and anti‐beta actin (ab8226; Abcam). Secondary antibodies were peroxidase anti‐mouse IgG (H + L) (PI‐2000; Vector Laboratories) and peroxidase anti‐rabbit IgG (H + L) (PI‐1000; Vector Laboratories). Bands were visualized using the myECL imager (Thermo Fisher Scientific).
Peroxidase anti mouse igg h l
Manufactured by Vector Laboratories
Peroxidase anti‐mouse IgG (H + L) is a secondary antibody that recognizes both the heavy and light chains of mouse immunoglobulin G (IgG). It is conjugated with horseradish peroxidase, an enzyme that can be used for detection in various immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Gradient polyacrylamide gel, by Bio-Rad (1 mentions)
Antibeta actin ab8226, by Abcam (1 mentions)
Peroxidase anti rabbit igg h l, by Vector Laboratories (1 mentions)
Myecl imager, by Thermo Fisher Scientific (1 mentions)
Caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
A21236, by Thermo Fisher Scientific (1 mentions)
Pi 2000, by Vector Laboratories (1 mentions)
A11031, by Thermo Fisher Scientific (1 mentions)
Hrp goat anti rabbit igg antibody, by Vector Laboratories (1 mentions)
F1804, by Merck Group (1 mentions)
2 protocols using peroxidase anti mouse igg h l
1
Apoptosis Detection by Western Blot
2
Western Blot and Immunofluorescence Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For Western blots, the
following primary antibodies were diluted in BSA (5%) in 1× PBS
buffer: monoclonal ANTI-FLAG M2 antibody produced in mouse (1:1000,
F1804, Merck); anti-β-actin antibody, mouse monoclonal (1:2000,
A1978, Merck). HRP goat antirabbit IgG antibody (peroxidase) (1:3000,
PI-1000, Vectorlabs), and peroxidase antimouse IgG (H+L) (affinity
purified) (1:3000, PI-2000, Vectorlabs) were used as secondary antibodies.
For IF assays, the following primary antibodies were diluted in
blocking buffer (FBS 3% in 1× PBS): the 5-hydroxymethylcytosine
(5hmC) antibody (pAb) (1:500, Active Motif cat#: 39769)
and monoclonal ANTI-FLAG M2 antibody produced in mouse (1:500, F1804,
Merck). Goat antimouse Alexa Fluor 568 (1:500, A-11031, Invitrogen)
and Goat antirabbit Alexa Fluor 647 (1:500, A-21236, Invitrogen) were
used as secondary antibodies.
following primary antibodies were diluted in BSA (5%) in 1× PBS
buffer: monoclonal ANTI-FLAG M2 antibody produced in mouse (1:1000,
F1804, Merck); anti-β-actin antibody, mouse monoclonal (1:2000,
A1978, Merck). HRP goat antirabbit IgG antibody (peroxidase) (1:3000,
PI-1000, Vectorlabs), and peroxidase antimouse IgG (H+L) (affinity
purified) (1:3000, PI-2000, Vectorlabs) were used as secondary antibodies.
For IF assays, the following primary antibodies were diluted in
blocking buffer (FBS 3% in 1× PBS): the 5-hydroxymethylcytosine
(5hmC) antibody (pAb) (1:500, Active Motif cat#: 39769)
and monoclonal ANTI-FLAG M2 antibody produced in mouse (1:500, F1804,
Merck). Goat antimouse Alexa Fluor 568 (1:500, A-11031, Invitrogen)
and Goat antirabbit Alexa Fluor 647 (1:500, A-21236, Invitrogen) were
used as secondary antibodies.
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