Commercially available antibodies used in this study are: ROR1, UHRF1 (#4102, #12387, Cell Signaling, Danvers MA), ROR1 (#AF2000, R&D Systems, Minneapolis MN), UHRF1 (#612264, BD Biosciences, San Jose CA), goat IgG serum (#005-000-003, Jackson ImmunoResearch, West Grove PA), mouse IgG serum (#sc-2025, Santa Cruz Biotechnology, Santa Cruz CA), actin, GAPDH, SP1 (#AM4300, #07-645, Millipore, Burlington MA), V5 (#46-0705, Invitrogen, Carlsbad CA), and ubiquitin (#A-104, Boston Biochem, Cambridge MA). The ROR1 antibody has been validated by the manufacturer and we have also tested this antibody in multiple cell lines to correlate immunoblot data with cell lines known to express or not express ROR1 at the transcript level (Supplementary Figure 5 ). Commercially available inhibitors used in this study are: Velcade (Bortezomib), dasatinib (SelleckChem, Houston TX), anisomycin (Cayman Chemical, Ann Arbor MI), naphthazarin, thymoquinone (Sigma, St. Louis MO), and flavopiridol (LC Laboratories, Woburn MA).
Uhrf1
Manufactured by BD
Sourced in United States
UHRF1 is a specialized lab equipment product offered by BD. It is a protein involved in the regulation of DNA methylation, a key epigenetic mechanism that plays a crucial role in various cellular processes. The core function of UHRF1 is to recognize and bind to hemimethylated DNA, facilitating the recruitment of DNA methyltransferases, which are responsible for maintaining DNA methylation patterns.
Automatically generated - may contain errors
Lab products found in correlation
Anisomycin, by Cayman Chemical (2 mentions)
Naphthazarin, by Merck Group (2 mentions)
Dasatinib, by Selleck Chemicals (2 mentions)
Actin, by Merck Group (2 mentions)
Ubiquitin, by R&D Systems (2 mentions)
Uhrf1, by Cell Signaling Technology (2 mentions)
Goat igg serum, by Jackson ImmunoResearch (2 mentions)
Mouse igg serum, by Santa Cruz Biotechnology (2 mentions)
Thymoquinone, by Merck Group (2 mentions)
Gapdh, by Merck Group (2 mentions)
Flavopiridol, by LC Laboratories (2 mentions)
Hdac1, by Thermo Fisher Scientific (2 mentions)
Dnmt1, by Thermo Fisher Scientific (2 mentions)
α4 integrin, by Thermo Fisher Scientific (2 mentions)
E cadherin, by Fortis Life Sciences (2 mentions)
P120 catenin, by Fortis Life Sciences (2 mentions)
Marveld2, by Merck Group (2 mentions)
β actin, by Thermo Fisher Scientific (2 mentions)
β catenin, by Fortis Life Sciences (2 mentions)
Epcam, by Thermo Fisher Scientific (2 mentions)
7 protocols using uhrf1
1
Antibody Validation and Inhibitor Usage
2
Antibody Validation and Inhibitor Usage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Commercially available antibodies used in this study are: ROR1, UHRF1 (#4102, #12387, Cell Signaling, Danvers MA), ROR1 (#AF2000, R&D Systems, Minneapolis MN), UHRF1 (#612264, BD Biosciences, San Jose CA), goat IgG serum (#005-000-003, Jackson ImmunoResearch, West Grove PA), mouse IgG serum (#sc-2025, Santa Cruz Biotechnology, Santa Cruz CA), actin, GAPDH, SP1 (#AM4300, #07-645, Millipore, Burlington MA), V5 (#46-0705, Invitrogen, Carlsbad CA), and ubiquitin (#A-104, Boston Biochem, Cambridge MA). The ROR1 antibody has been validated by the manufacturer and we have also tested this antibody in multiple cell lines to correlate immunoblot data with cell lines known to express or not express ROR1 at the transcript level (Supplementary Figure 5 ). Commercially available inhibitors used in this study are: Velcade (Bortezomib), dasatinib (SelleckChem, Houston TX), anisomycin (Cayman Chemical, Ann Arbor MI), naphthazarin, thymoquinone (Sigma, St. Louis MO), and flavopiridol (LC Laboratories, Woburn MA).
3
Biochemical Characterization of Necroptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TNF-α recombinant protein was generated as previously described.1 (link) z-VAD was from Bachem (Babendorf, Switzerland). Necrostatin-1 was purchased from Alexis Biochemicals (San Diego, CA, USA). The following antibodies were used for western blot analysis: RIP1 (BD Biosciences, NJ, USA, 610458), phospho-MLKL (Abcam, Cambridge, UK, 187091), caspase-8 (Cell Signaling, Danfoss, MA, USA, 9746), CYLD (Cell Signaling, 437700), mouse RIP3 (Prosci (San Diego, CA, USA), 2283), Sp1 (Santa Cruz, CA, USA, 14027), UHRF1 (BD Biosciences, 612264), Dnmt1 (Abcam, 13537), Flag (Sigma, St. Louis, MO, USA, A8592) and β-actin (Sigma, A2066). The following antibodies were used for flow cytometry analysis: CD11b (Biolegend, San Diego, CA, USA, 101212), CD45 (BD, 563891), GR-1 (BD Biosciences RB6-8C5), F4/80 (Biolegend, 123113).
4
Antibody-based Protein Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used in this study: UHRF1 (#612264, dilution: 1:1,000, BD Science) for western and IP and (H-8, dilution: 1:1000, Santa Cruz) for IF. BRCA1 (D-9, dilution: 1:100, Santa Cruz) for IF and (C-20, 1:200, Santa Cruz) for western blot and IP. RIF1 (A300-569A, dilution: 1:1,000, Bethyl Laborataries), HA (H9658, dilution: 1:1,000, Sigma), FLAG (F3165, dilution: 1:1,000, sigma), ubiquitin (SC-8017, dilution: 1:500, Santa Cruz), γ-H2AX (05-636, dilution: 1:500, Millipore), RPA (ab2175, dilution: 1:200, Abcam), RAD51(GTX70230, dilution: 1:200, GeneTex), 53BP1(NB100-304, dilution: 1:1,000, Novus), Phospho-(Ser) CDKs substrate antibody (9477s, dilution: 1:500, CST), cyclin A antibody (SC751, dilution: 1:200, Santa Cruz). CtIP (61141, dilution: 1:500, Active Motif). UHRF1, BRCA1 and RIF1 cDNAs were subcloned into the Flag-tagged vector (pIRES2-EGFP) HA-tagged vector (pCDNA3.1-HA or pCMV-HA). All mutants were generated by site-directed mutagenesis and confirmed by sequencing.
5
Antibody Immunoblotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies were obtained commercially from various sources – 1) Cell Signaling and Technology -Lamin A/C ( #4777); EpCAM (#2626S); DNMT1 (#5032P); HDAC1 (#5356P); G9a (3306S); Snail1 (3879P); α4-Integrin (#8440S); β-actin (#4970S); Di, Tri-methyl histone H3 (Lys9) (#5327) 2) Thermoscientific – UHRF1 (PA5-27969) 3) BD Biosciences – E-cadherin (610181); p120-catenin (610133); β-catenin (610154) 4) Bethyl laboratories – MARVELD2 (A301-505A) and 5) Sigma/Aldrich – β-tubulin (T0198).
6
Antibody Immunoblotting Protocol
7
Chromatin Immunoprecipitation and qRT-PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ChIP was performed by using SimpleChIP Enzymatic Chromatin IP kit (Cell Signaling Technology) according to the manufacturer's instructions. The antibodies used for ChIP are as follows: UHRF1 (612264; BD Biosciences, Franklin Lakes, NJ, USA); total histone H3 (4620), HDAC1 (34589), and HDAC2 (57156) from Cell Signaling Technology; acetyl‐histone H3 (06‐599) and acetyl‐histone H4 (06‐866) from Millipore. ChIP DNA was analyzed by PCR along with input DNA and quantified as % input for each target before calculation of fold changes or ratios of the signals among different experimental groups. The qRT‐PCR was performed by analyzing samples in triplicate using at least three independent sets of cDNA. The results were normalized by the expression level of actin as an internal control. The primer sequences used for the qRT‐PCR and ChIP assays are listed in supplementary data (Table S1 ).
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!