The antibodies used in this study are as follows: for ChIP-seq, RPB1 NTD (Cell Signaling Technology, 14958, lot #1), RPB1 Ser2 (Abcam, 5095, lot #GR3225147-1), H3K36me3 (Abcam, 9050, lot #GR3257952-1), H3K36me2 (Abcam, 9049, lot #GR3236147-1), CSTF64 (Bethyl Laboratories, A301-92A, lot #A301-092A-2), and CPSF73 (Bethyl Laboratories, A301-019A, lot #A301-091A-1); for Western blot, INTS11 (Atlas Antibodies, HPA029025, lot #A107128), lamin A (Active Motif, 39961, lot #33310001), TFIIB (Cell Signaling Technology, 4169s, lot #1), CPSF100 (Bethyl Laboratories, A301-583A-M, lot #A301-583A-M-1), CSTF50 (Bethyl Laboratories, A301-250A-M, lot #A301-250A-M-3), CSTF64 (Bethyl Laboratories, A301-092A-M, lot #A301-092A-M-2), CPSF73 (Bethyl Laboratories, A301-091A-M, lot #A301-091A-M-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab8245, lot #GR3317834-1).
Cstf 64
Manufactured by Fortis Life Sciences
The CSTF-64 is a laboratory instrument designed for the analysis of cellular samples. It is a centrifugal cell separating and testing system that can be used to isolate and purify various cell types from complex biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Cstf 77, by Fortis Life Sciences (2 mentions)
Cfim 68, by Fortis Life Sciences (2 mentions)
Cpeb1, by Cell Signaling Technology (2 mentions)
β actin, by Merck Group (2 mentions)
Bca assessment, by Thermo Fisher Scientific (2 mentions)
Cpsf73, by Fortis Life Sciences (2 mentions)
Wdr33, by Fortis Life Sciences (1 mentions)
H3k36me2, by Abcam (1 mentions)
Rpb1 ntd, by Cell Signaling Technology (1 mentions)
Tfiib, by Cell Signaling Technology (1 mentions)
H3k36me3, by Abcam (1 mentions)
Dab chromogen, by Roche (1 mentions)
Envision hrp conjugated secondary antibody, by Agilent Technologies (1 mentions)
Tubulin antibody, by Merck Group (1 mentions)
Ptbp1, by Abcam (1 mentions)
6 protocols using cstf 64
1
Antibody Validation for ChIP-seq and Western Blot
2
Multimodal Sequencing Techniques for Comprehensive Transcriptome Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3′-Seq was carried out using QuantSeq Rev 3ʹ mRNA sequencing library prep kit (Lexogen). Downstream sequence analysis was carried out using our recently published pipeline QuantifyPoly(A) (25 ).
ChIP-Seq libraries were prepared using the ChIP-IT Express Enzymatic Shearing (Active Motif) and ChIP-Seq library preparation (Vazyme) kits. Primary antibodies were purchased from Abcam or Santa Cruz Biotechnology. These libraries were sequenced on the NovaSeq platform, and all ChIP-Seq data processing and analysis were performed according to the ENCODE ChIP-Seq pipeline (https://www.encodeproject.org/data-standards/chip-seq/ ).
iCLIP-Seq library preparation and bioinformatics analysis were conducted as previously described (30 (link), 41 (link)). The primary antibodies (WDR33 [A301–152A] and CstF64 [A301–092A]) were purchased from Bethyl.
ChIP-Seq libraries were prepared using the ChIP-IT Express Enzymatic Shearing (Active Motif) and ChIP-Seq library preparation (Vazyme) kits. Primary antibodies were purchased from Abcam or Santa Cruz Biotechnology. These libraries were sequenced on the NovaSeq platform, and all ChIP-Seq data processing and analysis were performed according to the ENCODE ChIP-Seq pipeline (
iCLIP-Seq library preparation and bioinformatics analysis were conducted as previously described (30 (link), 41 (link)). The primary antibodies (WDR33 [A301–152A] and CstF64 [A301–092A]) were purchased from Bethyl.
3
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole cell lysates were prepared from −80°C-stored cell
pellets with RIPA lysis buffer, on ice. Following brief sonication, lysates were
clarified by 12,000 × g centrifugation for 10–15 min. Samples
were loaded according to total protein content, determined by BCA assessment
(Thermo Pierce), and within each cell type the amounts were normalized to the
first mock-infected sample (corresponding to ~100,000 cells total per
lane). Antibodies and dilutions used: CSTF-64 (A301-092A), CSTF-77 (A301-096A),
and CFIm-68 (A301-356A): Bethyl Laboratories, 1:2000. CPEB1: Cell
Signaling Technologies (13583), 1:1000. β-actin: Sigma-Aldrich (clone
AC-15, A1978), 1:10,000. Ms anti IE (CH160): Virusys, 1:5000. Ms anti UL57:
Virusys, 1:1000. Ms anti UL99(pp28): Virusys, 1:1000. Ms anti UL83(pp65)
Virusys, 1:5000.
pellets with RIPA lysis buffer, on ice. Following brief sonication, lysates were
clarified by 12,000 × g centrifugation for 10–15 min. Samples
were loaded according to total protein content, determined by BCA assessment
(Thermo Pierce), and within each cell type the amounts were normalized to the
first mock-infected sample (corresponding to ~100,000 cells total per
lane). Antibodies and dilutions used: CSTF-64 (A301-092A), CSTF-77 (A301-096A),
and CFIm-68 (A301-356A): Bethyl Laboratories, 1:2000. CPEB1: Cell
Signaling Technologies (13583), 1:1000. β-actin: Sigma-Aldrich (clone
AC-15, A1978), 1:10,000. Ms anti IE (CH160): Virusys, 1:5000. Ms anti UL57:
Virusys, 1:1000. Ms anti UL99(pp28): Virusys, 1:1000. Ms anti UL83(pp65)
Virusys, 1:5000.
4
Western Blot Analysis of Protein Expression
5
Immunohistochemical Analysis of Wound Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue slices (4 μm) from paraffin-embedded blocks containing wounds were de-paraffinized and rehydrated with a graded series of alcohols. Slides were subjected to heat-induced antigen retrieval with pH 6.0 citrate buffer. Slides were treated with primary antibodies against Ki-67 (Abcam, catalog no. ab16667, dilution 1:150), histone H4 (EMD Millipore, 05-858, 1:2000), CstF-64 (Bethyl Laboratories, IHC-00221, 1:1000), CPSF73 (Bethyl, A301-090A, 1:200) or CFIm25 (Sigma, AV40695, 1:200), followed by EnVision+ HRP-conjugated secondary antibody (Dako) and DAB chromogen (Roche) visualization. Slides were counterstained with hematoxylin and imaged with a Zeiss AXIO Imager.D2 microscope.
6
Antibody Sources for Pol II Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pol II antibodies CMA601, CMA602, and CMA603 were generated by HK (Stasevich et al., 2014 (link)). 8WG16, Aly, and PTBP1 antibodies were purchased from Abcam. CPSF73, CstF-64, and CstF-64 tau antibodies were purchased from Bethyl laboratories. Tubulin antibody was purchased from Sigma. Xrn2 antibody was provided by Dr. N. Gromak.
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!