Unless otherwise stated the following primary antibodies were used for western immunoblotting, diluted in 5% w/v BSA, 0.1% Tween-20 TBS and incubated at 4 °C overnight with gentle agitation; anti-ERCC1 antibody 1:100 (Santa Cruz Biotechnology, sc-1708), anti-FANCA 1:1,000 (Cell Signaling Technology, D1L2Z), anti-PLZF 1:100 (Santa Cruz Biotechnology, sc-28319), anti-Histone H3 1:5,000 (Abcam, 1791), anti-β-ACTIN 1:3,000 (Abcam, 8227), anti-VINCULLIN 1:2,000 (Abcam, 129002). The following secondary antibodies were diluted in 5% w/v BSA, 0.1% Tween-20 TBS and incubated for 1 hour at room temperature with gentle agitation; Swine anti-rabbit or anti-mouse HRP-conjugated immunoglobulins 1:2,000 (Dako). To demonstrate the absence of ERCC1 in Ercc1-mutant MEFs derived from Ercc1tm1a(KOMP)Wtsi embryos anti-ERCC1 1:1,000 (Cell Signaling Technology, D6G6) was employed, following the same western immunoblotting protocol described previously.
Anti ercc1
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-ERCC1 is a primary antibody that recognizes the ERCC1 protein, which is involved in the nucleotide excision repair pathway. This antibody can be used for applications such as Western blotting and immunohistochemistry to detect and study the ERCC1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti plzf, by Santa Cruz Biotechnology (2 mentions)
Anti ercc1 antibody, by Santa Cruz Biotechnology (2 mentions)
Anti fanca, by Cell Signaling Technology (2 mentions)
Anti vincullin, by Abcam (2 mentions)
Swine anti rabbit or anti mouse hrp conjugated immunoglobulins, by Agilent Technologies (2 mentions)
Anti histone h3, by Abcam (2 mentions)
Anti β actin, by Abcam (2 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (2 mentions)
Anti atm, by Cell Signaling Technology (2 mentions)
Anti ku70, by Cell Signaling Technology (2 mentions)
Caspase 3, by Cell Signaling Technology (2 mentions)
Anti dna pkcs, by Cell Signaling Technology (2 mentions)
Anti ku80, by Cell Signaling Technology (2 mentions)
Anti p95 nbs1, by Cell Signaling Technology (2 mentions)
Anti γh2ax, by Merck Group (2 mentions)
Anti cleaved parp, by Cell Signaling Technology (2 mentions)
Anti gapdh, by Santa Cruz Biotechnology (2 mentions)
Ab3594, by Abcam (1 mentions)
Ab2886, by Abcam (1 mentions)
Ab2621, by Abcam (1 mentions)
8 protocols using anti ercc1
1
Western Blotting Antibody Validation
2
Comprehensive Western Blotting Antibody Panel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies used in Western blotting were purchased from Cell Signaling Technology (Danvers, MA, USA): anti-HA-tag (6E2) (1:1000, #2367, Cell Signaling Technology), anti-p53 (7F5) (1:1000, #2527, Cell Signaling Technology), anti-DDB-1 (D4C8) (1:1000, #6998, Cell Signaling Technology), anti-DDB-2 (D4C4) (1:1000, #5416, Cell Signaling Technology) anti-ERCC1 (1:1000, #3885, Cell Signaling Technology), and anti-XPC (D1M5Y) (1:1000, #14768, Cell Signaling Technology). The Polyclonal anti-KMT4/DOT1L (1:2000, ab72454, Abcam) antibody, anti-XPA (1:2000, ab85914, Abcam), polyclonal di-methylated H3K79 antibody (1:1000, ab3594, Abcam), polyclonal to mono-methylated H3K79 antibody (1:500, ab2886, Abcam), and polyclonal tri-methylated H3K79 antibody (1:1000, ab2621, Abcam) were purchased from Abcam. TFIIH p62 Antibody (1:500, Q-19) was purchased from Santa Cruz Biotechnologies, Inc. Other antibodies include monoclonal Anti-β-Actin-Peroxidase antibody (1:25000, AC-15, Sigma Aldrich), monoclonal ANTI-FLAG® M2 antibody (1:1000, Sigma Aldrich), and anti-H3 (1:500, 865R2, Thermo Fisher Scientific Inc.). Most important uncropped blot images are shown in Supplementary Fig. 11 .
3
Quantitative Protein Analysis in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were obtained from tissue lysates with the RNA/Protein purification Kit (NorBiotek Canada, Thorold, ON, Canada). Additionally, protein extract from cell line were obtained by the RIPA 1× buffer supplemented with Complete® inhibitors cocktail (Roche Applied Science, Mannheim, Germany). Proteins were separated by 10% SDS-PAGE, blotted onto nitrocellulose membranes, and blocked. The membranes were then exposed to mouse anti-Phospho STAT6 (1:1000) or anti-Total-STAT6 (1:1000), anti-SNAI1, anti-ERCC1, or anti-phospho-ERK1/2 and anti-Total ERK 1/2 antibodies (all from Cell Signaling Technology, Danvers, MA, USA). The anti-β-actin monoclonal antibody (Santa Cruz Biotechnology, St. Cruz, CA, USA) was utilized to detect cell actin as the protein load control. Horseradish peroxidase (HRP)-tagged secondary anti-rabbit or anti-mouse antibodies (1:5000) (Jackson Immunoresearch, West Grove, PA, USA) were used for chemiluminescent detection with the ImmobilonTM Western Chemiluminescent HRP Substrate kit (Millipore, Burlington, MA, USA). Chemiluminescence signals were recorded on Alliance Q9 UVITEC imaging device (Cambridge, UK) and densitometric analyses were performed with ImageJ software.
4
DNA Damage Response Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-p53 and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-cleaved PARP, caspase-3, caspase-9, cleaved caspase-3, cleaved caspase-9, anti-LC3, anti-ATM, anti-p95/NBS1, anti-Ku70, anti-Ku80, anti-DNA-PKcs, and anti-ERCC1 were purchased from Cell Signaling Technology (Danvers, MA, USA), and anti-γ-H2AX was obtained from Millipore (Billerica, MA, USA).
5
Molecular Targets in Cancer Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-NF-κB, anti-Bcl-2, anti-cyclin A, anti-cyclin B, anti-cyclin E, anti-phospho-Raf-1, anti-Akt, anti-Snail, and anti-β-actin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-cleaved PARP, caspase-3, anti-ATM, anti-ATR, anti-Rad50, anti-p95/NBS1, anti-Rad52, anti-MRE11, anti-Ku70, anti-Ku80, anti-DNA-PKcs, anti-ERCC1, anti-Rad51, anti-Ras, anti-MEK1/2, anti-phospho-MEK1/2, anti-Erk1/2, anti-phospho-Erk1/2, anti-phospho-Akt, anti-vimentin, anti-E-cadherin, and anti-MMP9 were purchased from Cell Signaling Technology (Danvers, MA, USA), and anti-γ-H2AX was obtained from Millipore (Billerica, MA, USA). ZOL was purchased from Sigma-Aldrich (St. Louis, MO, USA). For in vitro experiments, ZOL was dissolved in PBS to make a 2 mmol/L stock solution and stored at −20°C.
6
HeLa Cell Culture and Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cervical carcinoma cell line, HeLa, was cultured in Dulbecco’s Modified Eagle Medium (DMEM) (GIBCO, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA) in glass Petri dishes at 37 °C in a 5% CO2 incubator.
Anti-dCK antibody was purchased from Abcam Inc (Cambridge, MA, USA). Anti-MAPLC3, anti-P-mTOR, anti-P-Akt, anti-P-P70S6K, anti-caspase3, anti-Bcl-2, anti-H2AX and anti-ERCC1 antibodies were purchased from Cell Signaling. Anti-GAPDH was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The peroxidase-conjugated anti-mouse IgG and peroxidase-conjugated anti-rabbit IgG were purchased at Santa Cruz. fetal bovine serum (FBS), and Cell Counting Kit-8 (CCK-8) was purchased from Dojin Laboratories (Kumamoto, Japan), 3-Methyladenine (3-MA), NH4Cl, Necrostatin-1, Ferrostatin-1, Rapamycin, Spautin-1 and monodansylcadaverine (MDC) were purchased from Sigma Chemical (St. Louis, MO, USA), ZVAD-FMK was purchased from Selleckchem (Houston, TX, USA), and pSUPER retroviral vector was obtained from OligoEngine (Seattle, WA, USA).
Anti-dCK antibody was purchased from Abcam Inc (Cambridge, MA, USA). Anti-MAPLC3, anti-P-mTOR, anti-P-Akt, anti-P-P70S6K, anti-caspase3, anti-Bcl-2, anti-H2AX and anti-ERCC1 antibodies were purchased from Cell Signaling. Anti-GAPDH was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The peroxidase-conjugated anti-mouse IgG and peroxidase-conjugated anti-rabbit IgG were purchased at Santa Cruz. fetal bovine serum (FBS), and Cell Counting Kit-8 (CCK-8) was purchased from Dojin Laboratories (Kumamoto, Japan), 3-Methyladenine (3-MA), NH4Cl, Necrostatin-1, Ferrostatin-1, Rapamycin, Spautin-1 and monodansylcadaverine (MDC) were purchased from Sigma Chemical (St. Louis, MO, USA), ZVAD-FMK was purchased from Selleckchem (Houston, TX, USA), and pSUPER retroviral vector was obtained from OligoEngine (Seattle, WA, USA).
7
Analyzing Cellular Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot was performed as described previously. Samples were collected directly in 1X NuPAGE LDS sample buffer with 1X sample-reducing buffer (Invitrogen) and denatured at 95°C for 5 min followed by a centrifugation at 13200 rpm for 5 min. The supernatant was electrophoresed on a 4–12% Tris-HCl gel and transferred to nitrocellulose membranes (Invitrogen). After blocking with Superblock T20 blocking buffer (Thermo Scientific), the membranes were incubated with a primary antibody overnight at 4°C and then with a secondary antibody conjugated with alkaline phosphatase for 1 hour each at room temperature; the signal was detected using a chemiluminescence method. The following primary antibodies were used: anti-p53 (Cell Signaling, 1:1000); anti-phospho-p53 (S15; Cell Signaling, 1:1000); anti-GAPDH (Santa Cruz, 1:10000); anti-ERCC1 (Cell Signaling, 1:1000); and anti-PARP (Cell Signaling, 1:1000).
8
Western Blotting Antibody Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unless otherwise stated the following primary antibodies were used for western immunoblotting, diluted in 5% w/v BSA, 0.1% Tween-20 TBS and incubated at 4 °C overnight with gentle agitation; anti-ERCC1 antibody 1:100 (Santa Cruz Biotechnology, sc-1708), anti-FANCA 1:1,000 (Cell Signaling Technology, D1L2Z), anti-PLZF 1:100 (Santa Cruz Biotechnology, sc-28319), anti-Histone H3 1:5,000 (Abcam, 1791), anti-β-ACTIN 1:3,000 (Abcam, 8227), anti-VINCULLIN 1:2,000 (Abcam, 129002). The following secondary antibodies were diluted in 5% w/v BSA, 0.1% Tween-20 TBS and incubated for 1 hour at room temperature with gentle agitation; Swine anti-rabbit or anti-mouse HRP-conjugated immunoglobulins 1:2,000 (Dako). To demonstrate the absence of ERCC1 in Ercc1-mutant MEFs derived from Ercc1tm1a(KOMP)Wtsi embryos anti-ERCC1 1:1,000 (Cell Signaling Technology, D6G6) was employed, following the same western immunoblotting protocol described previously.
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!