Journal: STAR Protocols

Article Title: Protocol for Disome Profiling to Survey Ribosome Collision in Humans and Zebrafish

doi: 10.1016/j.xpro.2020.100168

Figure Lengend Snippet:

Article Snippet: SAPPHIRE FILTER TIP, 300 μL XL, LOW RETENTION, NATURAL, STERILE, 96 PCS./RACK , Greiner Bio-One , Cat# 738265.

Techniques: Recombinant, Reverse Transcription, Magnetic Beads, Staining, Sterility, Transferring, Electrophoresis, Blocking Assay, MicroChIP Assay, Spectrophotometry

Journal:

Article Title: The Papillomavirus E7 Oncoprotein Is Ubiquitinated by UbcH7 and Cullin 1- and Skp2-Containing E3 Ligase

doi: 10.1128/JVI.78.10.5338-5346.2004

Figure Lengend Snippet: Ubiquitination of E7 in cell-free lysates. (A and B) Purified His-E7 protein (0.5 μg) was ubiquitinated by using HeLa cell cytosolic extracts (50 μg) in a reaction mixture containing ubiquitin and ATP as described in Materials and Methods. The E7-ubiquitin conjugates were purified with an Ni affinity column, divided into two parts, and resolved by SDS-10% PAGE. One part (A) was probed with an E7 antibody and the other part (B) was probed with a ubiquitin (Ub) antibody in Western blot assays. (C) 35S-labeled nontagged HPV16 E7 made in rabbit reticulocyte lysates was ubiquitinated with HeLa cell S-100 (50 μg) in the presence of wild-type ubiquitin or ubiquitin-KR. The ubiquitinated proteins were immunoprecipitated (IP) with the E7 antibody, resolved by SDS-10% PAGE, and detected by fluorography.

Article Snippet: The purified recombinant yeast ubiquitin-activating enzyme E1, the ubiquitin-conjugating enzyme E2, and ubiquitin aldehyde were from Boston Biochem, Cambridge, Mass.

Techniques: Purification, Affinity Column, Western Blot, Labeling, Immunoprecipitation

Journal:

Article Title: The Papillomavirus E7 Oncoprotein Is Ubiquitinated by UbcH7 and Cullin 1- and Skp2-Containing E3 Ligase

doi: 10.1128/JVI.78.10.5338-5346.2004

Figure Lengend Snippet: HeLa cell FrI and FrIIA together promote the formation of E7-ubiquitin conjugates. Ubiquitination reactions were carried out as described in Materials and Methods with His-E7 (0.3 μg), ubiquitin (5 μg), recombinant E1 (30 ng) and, when indicated, HeLa cell Fr1 (25 μg), FrIIA (25 μg), FrIIB (25 μg), or S-100 (30 μg). The E7-ubiquitin (Ub-E7) conjugates were purified with an Ni affinity column, resolved by SDS-10% PAGE, and probed with a ubiquitin antibody in Western blot assays.

Article Snippet: The purified recombinant yeast ubiquitin-activating enzyme E1, the ubiquitin-conjugating enzyme E2, and ubiquitin aldehyde were from Boston Biochem, Cambridge, Mass.

Techniques: Recombinant, Purification, Affinity Column, Western Blot

Journal:

Article Title: The Papillomavirus E7 Oncoprotein Is Ubiquitinated by UbcH7 and Cullin 1- and Skp2-Containing E3 Ligase

doi: 10.1128/JVI.78.10.5338-5346.2004

Figure Lengend Snippet: UbcH7 is involved in the conjugation of ubiquitin to E7. (A) His-E7 protein (0.65 μg) was ubiquitinated by using HeLa cell FrIIA (50 μg), recombinant E1 protein (30 ng), and the indicated recombinant E2 protein (300 ng) in ubiquitination assays as described in Materials and Methods. The E7-ubiquitin conjugates were purified with an Ni affinity column, resolved by SDS-11% PAGE, and probed with an E7 antibody. (B) 35S-labeled E7 made in rabbit reticulocyte lysates was ubiquitinated with recombinant E1 protein (30 ng), HeLa cell FrIIA (25 μg), and the indicated recombinant E2 protein (300 ng). The reaction mixtures were resolved by SDS-10% PAGE, and the proteins were developed by fluorography. (C) Proteins (200 μg) from HeLa cell S-100, FrI, FrIIA, or FrIIB were resolved by SDS-12% PAGE and probed with a UbcH7 antibody in Western blot assays.

Article Snippet: The purified recombinant yeast ubiquitin-activating enzyme E1, the ubiquitin-conjugating enzyme E2, and ubiquitin aldehyde were from Boston Biochem, Cambridge, Mass.

Techniques: Conjugation Assay, Recombinant, Purification, Affinity Column, Labeling, Western Blot

Journal:

Article Title: The Papillomavirus E7 Oncoprotein Is Ubiquitinated by UbcH7 and Cullin 1- and Skp2-Containing E3 Ligase

doi: 10.1128/JVI.78.10.5338-5346.2004

Figure Lengend Snippet: E7 is ubiquitinated by the Cul1 complex and UbcH7 in vitro. C33A cells were transfected with a V5 epitope-tagged Cul1 expression plasmid, and the transfected cell extracts (2 mg) were immunoprecipitated (IP) with a control antibody or a V5 antibody (V5-Ab) and protein G-Sepharose beads. After brief washing, the beads with the immune complexes were used to ubiquitinate His-E7 (0.15 μg) as described in Materials and Methods. After the reaction, the beads were separated from the supernatants (Sup) by centrifugation. Proteins associated with the beads and in the supernatants were resolved separately by SDS-10% PAGE and probed with E7 and ubiquitin (Ub) monoclonal antibodies. IgG, immunoglobulin G; E7-Ub, E7-ubiquitin.

Article Snippet: The purified recombinant yeast ubiquitin-activating enzyme E1, the ubiquitin-conjugating enzyme E2, and ubiquitin aldehyde were from Boston Biochem, Cambridge, Mass.

Techniques: In Vitro, Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Centrifugation

Journal:

Article Title: The Papillomavirus E7 Oncoprotein Is Ubiquitinated by UbcH7 and Cullin 1- and Skp2-Containing E3 Ligase

doi: 10.1128/JVI.78.10.5338-5346.2004

Figure Lengend Snippet: Ubiquitinated E7 is accumulated in the detergent-insoluble nuclear fraction of MG132-treated Caski cells. (A) Caski cells were treated with DMSO or MG132 (10 μM) for 5 h before being harvested. The cells were lysed and separated into detergent-soluble (S) and detergent-insoluble (In) extracts as described in Materials and Methods. Detergent-soluble (300 μg) and detergent-insoluble (300 μg) cell extracts were separated by SDS-10% PAGE and probed by immunoblotting with an E7 antibody. (B and C) Detergent-soluble (1.5 mg) and detergent-insoluble (0.41 mg) cell extracts were immunoprecipitated with an E7 polyclonal antibody. The immunoprecipitated proteins were collected with protein A-Sepharose beads, separated by SDS-10% PAGE, and probed with an E7 monoclonal antibody (B) or a ubiquitin antibody (C). IgG, immunoglobulin G; Ub-E7, ubiquitin-E7.

Article Snippet: The purified recombinant yeast ubiquitin-activating enzyme E1, the ubiquitin-conjugating enzyme E2, and ubiquitin aldehyde were from Boston Biochem, Cambridge, Mass.

Techniques: Western Blot, Immunoprecipitation

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The primers and TaqMan probes of IGF, ER genes used for RT-PCR

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques:

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The mRNA levels of IGF-1, IGF-1R, IGF-2, IGF-2R, IGFBP-3, ERα and ERβ in EAC, tumor-adjacent, and control groups

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Control

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The immunostaining of IGF-1R, IGF-2R, ERα and ERβ in EAC, atypical hyperplasia, and control groups

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Immunostaining, Control, Positive Control

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The relationship between the IGF and ER mRNA expression and clinico-pathological features of endometrioid adenocarcinoma

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Expressing

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The relationship between the IGF-1R, IGF2R, ERα and ERβ protein expression and clinico-pathological features of endometrioid adenocarcinoma

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Expressing

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: Correlation analysis of IGF and ER mRNA expression with clinico-pathological features of EAC

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Expressing

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: Correlation analysis of IGF-1R, IGF-2R, and ERα and ERβ protein expression with clinico-pathological features of EAC

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Expressing

Journal: BMC Cancer

Article Title: Insulin-like growth factors in endometrioid adenocarcinoma: Correlation with clinico-pathological features and estrogen receptor expression

doi: 10.1186/1471-2407-12-262

Figure Lengend Snippet: The analysis of the correlation between the mRNA expression of IGFs with ER subtypes in three groups

Article Snippet: Bio-engineering Co ., Ltd ., Wuhan , China), IGF-2R (polyclonal rabbit anti-human IGF-2R, 1:50 dilution, Wuhan Boster .

Techniques: Expressing, Control