Western immunoblotting was accomplished as previously described (25 (link),26 (link)) using the following primary antibodies: anti-caspase-9 (Assay Designs); anti-FLAG and anti-β-actin (Sigma); anti-p65, anti-NF-κB2, anti-IκBα, phospho-IκBα, anti-laminA/C, anti-α-tubulin, anti-NIK, anti-RIP1, anti-cIAP1, anti-cIAP2, anti-Apaf1, anti-myc (Cell Signaling Technology); anti-K48, anti-Ki-67, and K63-linkage specific ubiquitin, and anti-Ki-67 (Abcam). Secondary antibodies were horseradish peroxidase-conjugated anti-rabbit IgG and anti-mouse IgG antibodies (Cell Signaling Technology).
Anti apaf 1
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-Apaf-1 is a laboratory reagent used in research applications. It is an antibody that specifically binds to the Apaf-1 (Apoptotic Protease Activating Factor 1) protein, which is a key component of the apoptosis (programmed cell death) signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Anti tubulin, by Cell Signaling Technology (2 mentions)
Anti bax, by Cell Signaling Technology (2 mentions)
Anti fas, by Cell Signaling Technology (2 mentions)
Anti caspases 8 and 9, by Cell Signaling Technology (2 mentions)
Anti fadd, by Cell Signaling Technology (2 mentions)
Anti bak, by Cell Signaling Technology (2 mentions)
Anti bid, by Cell Signaling Technology (2 mentions)
Anti lamin a c, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated anti rabbit igg and anti mouse igg antibodies, by Cell Signaling Technology (1 mentions)
Anti myc, by Cell Signaling Technology (1 mentions)
Anti flag and anti β actin, by Merck Group (1 mentions)
Anti p65, by Merck Group (1 mentions)
Anti nik, by Cell Signaling Technology (1 mentions)
Anti rip1, by Cell Signaling Technology (1 mentions)
Anti ciap1, by Cell Signaling Technology (1 mentions)
Anti ciap2, by Cell Signaling Technology (1 mentions)
Anti α tubulin, by Cell Signaling Technology (1 mentions)
Anti ki67, by Abcam (1 mentions)
Phospho iκbα, by Cell Signaling Technology (1 mentions)
Anti iκbα, by Cell Signaling Technology (1 mentions)
7 protocols using anti apaf 1
1
Western Blot Antibody Analysis
2
Apoptosis Signaling Pathway Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-FasL, anti-caspase-8, anti-β-actin, anti-Bax, anti-Bad, anti-MMP-2 and anti-MMP-9 were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Anti-cleaved caspase-3, anti-cleaved poly(ADP-ribose) polymerase (PARP), anti-cleaved caspase-9, anti-Bcl-2, anti-Bcl-xL, anti-Bad, anti-Apaf-1, phospho-Erk1/2, total-Erk1/2, phospho-p38, total-p38, phospho-JNK and total JNK were purchased from Cell Signaling (Danvers, MA, USA). ERK chemical inhibitor (PD98059) and p38 chemical inhibitor were purchased from EMD Chemicals (Gibbstown, NJ, USA).
3
Apoptosis Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standard conditions were used as previously described [18 (link)]. All antibodies (e.g., anti-Caspases-8 and -9, anti-Apaf-1, anti-BID, anti-FADD, anti-FAS, anti-BAX, anti-BAK and anti-Tubulin) were purchased from Cell Signaling Technology (Danvers, MA, USA) and utilized according to the manufacturer’s protocol.
4
Apoptosis Pathway Regulation in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The reagents and kits used in this study included DMEM (Hyclone Laboratories, Inc., Logan, UT, USA), FBS (Wisent, Nanjing, China), trypsin, DMSO, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Beyotime Institute of Biotechnology, Nanjing, China), mitochondrial protein extraction kit (KeyGen Biotech Co., Ltd., Nanjing, China), and Annexin V-FITC apoptosis detection kit (Bender Medsystems, San Diego, CA, USA). Target antibodies, including anti-Apaf-1 and anti-cytochrome c (Cyto C) primary antibodies, were all purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). The secondary antibody used was horseradish peroxidase (HRP)-goat anti rabbit IgG (Cell Signaling Technology, Inc.). β-actin (Beyotime Institute of Biotechnology) was used as an internal standard. The enhanced chemiluminescence (ECL) kit and polyvinylidene fluoride (PVDF) membranes were both purchased from Pierce (Rockford, IL, USA) and EMD Millipore (Billerica, MA, USA), respectively.
5
Apoptosis Inhibitors and Assay Methods
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis inhibitors, Hoechst 33258, 4′-6-diamidino-2-phenylindole (DAPI), and cell-permeant inhibitors of Apaf-1 (NS3694), caspase-8 (Z-IETD-FMK), caspase-9 (Z-LEHD-FMK), and caspase-3 (Z-EDVD-FMK) were obtained from Sigma-Aldrich (Saint Louis, MO). Fetal calf serum was from Cansera International Inc. (Etobicoke, Canada). STS, gentamicin, penicillin, streptomycin, and cycloheximide were from Wako (Tokyo, Japan). Anti-Apaf-1, anti-caspase-9 and anti-caspase-3 antibodies were from Cell Signaling Technology (Danvers, MA). anti-caspase-9 antibodies were also purchased from Calbiochem (La Jolla, CA) and Abcam (Cambridge, UK). Caspase-9, -8 and -3 Colorimetric Activity Assay kits, and ApopTag Fluorescein kit for TUNEL assays were from Chemicon (Temecula, CA). Chlamydiaceae-specific fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody (Chlamydia-FA) was from Denka Seiken (Tokyo, Japan).
6
Investigating KCNJ15 and APAF-1 Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following chemicals were used: KCl (Sigma-Aldrich, P954), paraformaldehyde (Electron Microscopy Sciences, 15710). The following antibodies were used: anti-KCNJ15 (Sigma-Aldrich, HPA016702), anti-APAF-1 (Cell Signaling Technology, 5088), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (14C10) (Cell Signaling Technology, 2118), anti-β-actin (Cell Signaling Technology, 4967), anti-rabbit immunoglobulin G (IgG) horseradish peroxidase (HRP)-linked antibody (Cell Signaling Technology, 7074). ON-TARGET plus SMARTpool Human KCNJ15 (3772) small interfering RNAs (siRNAs) (L006245000005) and control siRNAs (1299001) were from Dharmacon and Integrated DNA Technologies, respectively.
7
Lipid Peroxidation Assay in A375 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standard conditions were used as previously described [18] . All antibodies (e.g., anti-Caspases-8 and -9, anti-Apaf-1, anti-BID, anti-FADD, anti-FAS, anti-BAX, anti-BAK and anti-Tubulin) were purchased from Cell Signaling Technology (Danvers, MA, USA) and utilized according to the manufacturer's protocol.
2.9. Determination of lipid peroxidation content A375 cells were plated in 100 mm dishes, cultured overnight and next day were treated with L-SK-4 (100 µM). After trypsinization, pellets were collected, re-suspended and sonicated before the TBARS Assay kit (Cambridge Bioscience Ltd, Cambridge, UK) was utilized for the determination of malondialdehyde (MDA) content according to the manufacture's protocol.
2.9. Determination of lipid peroxidation content A375 cells were plated in 100 mm dishes, cultured overnight and next day were treated with L-SK-4 (100 µM). After trypsinization, pellets were collected, re-suspended and sonicated before the TBARS Assay kit (Cambridge Bioscience Ltd, Cambridge, UK) was utilized for the determination of malondialdehyde (MDA) content according to the manufacture's protocol.
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!