rabbit polyclonal anti e1b 19k  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Name:
    Monoclonal ANTI FLAG M2 antibody
    Description:
    Anti Flag M2 antibody is used for the detection of Flag fusion proteins This monoclonal antibody is produced in mouse and recognizes the FLAG sequence at the N terminus Met N terminus and C terminus The antibody is also able to recognize FLAG at an internal site M2 unlike M1 antibody is not Calcium dependent F1804 is an affinity purified FLAG M2 antibody increasing sensitivity in most applications Method of purification Protein A
    Catalog Number:
    f3165
    Price:
    None
    Applications:
    Antibody is recommended for use in immunoblotting, immunoprecipitation, immunocytochemistry, immunofluorescence, ELISA, electron microscopy, flow cytometry and supershift assays.Browse additional application references in our FLAG(R) Literature portal.
    Buy from Supplier


    Structured Review

    Millipore rabbit polyclonal anti e1b 19k
    Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or <t>E1B</t> <t>19K.</t> Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).
    Anti Flag M2 antibody is used for the detection of Flag fusion proteins This monoclonal antibody is produced in mouse and recognizes the FLAG sequence at the N terminus Met N terminus and C terminus The antibody is also able to recognize FLAG at an internal site M2 unlike M1 antibody is not Calcium dependent F1804 is an affinity purified FLAG M2 antibody increasing sensitivity in most applications Method of purification Protein A
    https://www.bioz.com/result/rabbit polyclonal anti e1b 19k/product/Millipore
    Average 85 stars, based on 7213 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti e1b 19k - by Bioz Stars, 2020-09
    85/100 stars

    Images

    1) Product Images from "Hypoxia and defective apoptosis drive genomic instability and tumorigenesis"

    Article Title: Hypoxia and defective apoptosis drive genomic instability and tumorigenesis

    Journal: Genes & Development

    doi: 10.1101/gad.1204904

    Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).
    Figure Legend Snippet: Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).

    Techniques Used: Staining, Transformation Assay, Expressing, Injection, Derivative Assay, Immunohistochemistry, Mouse Assay

    Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.
    Figure Legend Snippet: Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.

    Techniques Used: Blocking Assay, Stable Transfection, Western Blot, Expressing, Clone Assay, Plasmid Preparation, Injection, Mouse Assay

    2) Product Images from "Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿ †"

    Article Title: Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿ †

    Journal: Journal of Virology

    doi: 10.1128/JVI.01797-09

    The pan-caspase inhibitor zVAD-FMK protects NS1-expressing and H-1PV-infected cells from undergoing apoptosis. 293-NS1 cells grown in medium supplemented (induced) or not supplemented (noninduced) with doxycycline, and HEK-293 cells either mock treated
    Figure Legend Snippet: The pan-caspase inhibitor zVAD-FMK protects NS1-expressing and H-1PV-infected cells from undergoing apoptosis. 293-NS1 cells grown in medium supplemented (induced) or not supplemented (noninduced) with doxycycline, and HEK-293 cells either mock treated

    Techniques Used: Expressing, Infection

    NS1 induces apoptosis in HEK-293 cells. (A) Cells grown in the presence (induced) or absence (noninduced) of doxycycline were harvested at the indicated times, fixed, labeled with propidium iodide, and analyzed by FACS. At least 15,000 events were analyzed
    Figure Legend Snippet: NS1 induces apoptosis in HEK-293 cells. (A) Cells grown in the presence (induced) or absence (noninduced) of doxycycline were harvested at the indicated times, fixed, labeled with propidium iodide, and analyzed by FACS. At least 15,000 events were analyzed

    Techniques Used: Labeling, FACS

    NS1 arrests the cell cycle at the G 2 /M stage. Parental and stably transfected 293-NS1 cells were cultured in medium supplemented (ind.) or not supplemented (non ind.) with doxycycline for 12 or 24 h and processed for Western blot analysis and FACS determination
    Figure Legend Snippet: NS1 arrests the cell cycle at the G 2 /M stage. Parental and stably transfected 293-NS1 cells were cultured in medium supplemented (ind.) or not supplemented (non ind.) with doxycycline for 12 or 24 h and processed for Western blot analysis and FACS determination

    Techniques Used: Stable Transfection, Transfection, Cell Culture, Western Blot, FACS

    H-1PV NS1 induces DNA damages via ROS production. (A) Cells were analyzed for the presence of DNA damages by immunofluorescence using an anti-γ-H2AX antibody at 24 h. NS1-expressing and H-1PV-infected cells display a specific nuclear staining
    Figure Legend Snippet: H-1PV NS1 induces DNA damages via ROS production. (A) Cells were analyzed for the presence of DNA damages by immunofluorescence using an anti-γ-H2AX antibody at 24 h. NS1-expressing and H-1PV-infected cells display a specific nuclear staining

    Techniques Used: Immunofluorescence, Expressing, Infection, Staining

    The 293-NS1 system allows the tightly controlled induction of H-1PV NS1 expression. The human T-REx Flp-In HEK-293 embryonic kidney cell line expressing the full-length H-1PV NS1 gene under the control of a doxycycline-inducible promoter (293-NS1) was
    Figure Legend Snippet: The 293-NS1 system allows the tightly controlled induction of H-1PV NS1 expression. The human T-REx Flp-In HEK-293 embryonic kidney cell line expressing the full-length H-1PV NS1 gene under the control of a doxycycline-inducible promoter (293-NS1) was

    Techniques Used: Expressing

    Tentative model of H-1PV cytotoxicity. After parvovirus infection, expression of NS1 triggers an oxidative stress with accumulation of intracellular ROS associated with DNA damage. As a consequence of DNA damage, cells accumulate in G 2 phase and then
    Figure Legend Snippet: Tentative model of H-1PV cytotoxicity. After parvovirus infection, expression of NS1 triggers an oxidative stress with accumulation of intracellular ROS associated with DNA damage. As a consequence of DNA damage, cells accumulate in G 2 phase and then

    Techniques Used: Infection, Expressing

    NS1 expression is associated with differential expression of cell cycle regulatory proteins. Doxycycline was added to the medium (1,000 ng/ml) of 293-NS1 cultures. Whole cellular lysates were prepared every 4 h and subjected to SDS-PAGE fractionation
    Figure Legend Snippet: NS1 expression is associated with differential expression of cell cycle regulatory proteins. Doxycycline was added to the medium (1,000 ng/ml) of 293-NS1 cultures. Whole cellular lysates were prepared every 4 h and subjected to SDS-PAGE fractionation

    Techniques Used: Expressing, SDS Page, Fractionation

    NS1 expression is sufficient to induce cell lysis. Cell lysis and viability were measured by LDH and MTT assays, respectively, in parental 293 and 293-NS1 cultures grown in the absence (−) or presence (+) of doxycycline (1,000 ng/ml) for
    Figure Legend Snippet: NS1 expression is sufficient to induce cell lysis. Cell lysis and viability were measured by LDH and MTT assays, respectively, in parental 293 and 293-NS1 cultures grown in the absence (−) or presence (+) of doxycycline (1,000 ng/ml) for

    Techniques Used: Expressing, Lysis, MTT Assay

    Involvement of ROS production in NS1-induced apoptosis. (A) The levels of ROS in control 293-NS1 cells (non ind., purple) or cells expressing NS1 in the presence (ind.+NAC, red) or absence (ind., green) of the antioxidant NAC (5 mM) were determined
    Figure Legend Snippet: Involvement of ROS production in NS1-induced apoptosis. (A) The levels of ROS in control 293-NS1 cells (non ind., purple) or cells expressing NS1 in the presence (ind.+NAC, red) or absence (ind., green) of the antioxidant NAC (5 mM) were determined

    Techniques Used: Expressing

    3) Product Images from "Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿ †"

    Article Title: Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿Through Its Nonstructural Protein NS1, Parvovirus H-1 Induces Apoptosis via Accumulation of Reactive Oxygen Species ▿ †

    Journal: Journal of Virology

    doi: 10.1128/JVI.01797-09

    The pan-caspase inhibitor zVAD-FMK protects NS1-expressing and H-1PV-infected cells from undergoing apoptosis. 293-NS1 cells grown in medium supplemented (induced) or not supplemented (noninduced) with doxycycline, and HEK-293 cells either mock treated
    Figure Legend Snippet: The pan-caspase inhibitor zVAD-FMK protects NS1-expressing and H-1PV-infected cells from undergoing apoptosis. 293-NS1 cells grown in medium supplemented (induced) or not supplemented (noninduced) with doxycycline, and HEK-293 cells either mock treated

    Techniques Used: Expressing, Infection

    NS1 induces apoptosis in HEK-293 cells. (A) Cells grown in the presence (induced) or absence (noninduced) of doxycycline were harvested at the indicated times, fixed, labeled with propidium iodide, and analyzed by FACS. At least 15,000 events were analyzed
    Figure Legend Snippet: NS1 induces apoptosis in HEK-293 cells. (A) Cells grown in the presence (induced) or absence (noninduced) of doxycycline were harvested at the indicated times, fixed, labeled with propidium iodide, and analyzed by FACS. At least 15,000 events were analyzed

    Techniques Used: Labeling, FACS

    NS1 arrests the cell cycle at the G 2 /M stage. Parental and stably transfected 293-NS1 cells were cultured in medium supplemented (ind.) or not supplemented (non ind.) with doxycycline for 12 or 24 h and processed for Western blot analysis and FACS determination
    Figure Legend Snippet: NS1 arrests the cell cycle at the G 2 /M stage. Parental and stably transfected 293-NS1 cells were cultured in medium supplemented (ind.) or not supplemented (non ind.) with doxycycline for 12 or 24 h and processed for Western blot analysis and FACS determination

    Techniques Used: Stable Transfection, Transfection, Cell Culture, Western Blot, FACS

    H-1PV NS1 induces DNA damages via ROS production. (A) Cells were analyzed for the presence of DNA damages by immunofluorescence using an anti-γ-H2AX antibody at 24 h. NS1-expressing and H-1PV-infected cells display a specific nuclear staining
    Figure Legend Snippet: H-1PV NS1 induces DNA damages via ROS production. (A) Cells were analyzed for the presence of DNA damages by immunofluorescence using an anti-γ-H2AX antibody at 24 h. NS1-expressing and H-1PV-infected cells display a specific nuclear staining

    Techniques Used: Immunofluorescence, Expressing, Infection, Staining

    The 293-NS1 system allows the tightly controlled induction of H-1PV NS1 expression. The human T-REx Flp-In HEK-293 embryonic kidney cell line expressing the full-length H-1PV NS1 gene under the control of a doxycycline-inducible promoter (293-NS1) was
    Figure Legend Snippet: The 293-NS1 system allows the tightly controlled induction of H-1PV NS1 expression. The human T-REx Flp-In HEK-293 embryonic kidney cell line expressing the full-length H-1PV NS1 gene under the control of a doxycycline-inducible promoter (293-NS1) was

    Techniques Used: Expressing

    Tentative model of H-1PV cytotoxicity. After parvovirus infection, expression of NS1 triggers an oxidative stress with accumulation of intracellular ROS associated with DNA damage. As a consequence of DNA damage, cells accumulate in G 2 phase and then
    Figure Legend Snippet: Tentative model of H-1PV cytotoxicity. After parvovirus infection, expression of NS1 triggers an oxidative stress with accumulation of intracellular ROS associated with DNA damage. As a consequence of DNA damage, cells accumulate in G 2 phase and then

    Techniques Used: Infection, Expressing

    NS1 expression is associated with differential expression of cell cycle regulatory proteins. Doxycycline was added to the medium (1,000 ng/ml) of 293-NS1 cultures. Whole cellular lysates were prepared every 4 h and subjected to SDS-PAGE fractionation
    Figure Legend Snippet: NS1 expression is associated with differential expression of cell cycle regulatory proteins. Doxycycline was added to the medium (1,000 ng/ml) of 293-NS1 cultures. Whole cellular lysates were prepared every 4 h and subjected to SDS-PAGE fractionation

    Techniques Used: Expressing, SDS Page, Fractionation

    NS1 expression is sufficient to induce cell lysis. Cell lysis and viability were measured by LDH and MTT assays, respectively, in parental 293 and 293-NS1 cultures grown in the absence (−) or presence (+) of doxycycline (1,000 ng/ml) for
    Figure Legend Snippet: NS1 expression is sufficient to induce cell lysis. Cell lysis and viability were measured by LDH and MTT assays, respectively, in parental 293 and 293-NS1 cultures grown in the absence (−) or presence (+) of doxycycline (1,000 ng/ml) for

    Techniques Used: Expressing, Lysis, MTT Assay

    Involvement of ROS production in NS1-induced apoptosis. (A) The levels of ROS in control 293-NS1 cells (non ind., purple) or cells expressing NS1 in the presence (ind.+NAC, red) or absence (ind., green) of the antioxidant NAC (5 mM) were determined
    Figure Legend Snippet: Involvement of ROS production in NS1-induced apoptosis. (A) The levels of ROS in control 293-NS1 cells (non ind., purple) or cells expressing NS1 in the presence (ind.+NAC, red) or absence (ind., green) of the antioxidant NAC (5 mM) were determined

    Techniques Used: Expressing

    4) Product Images from "Human Adenovirus Core Protein V Is Targeted by the Host SUMOylation Machinery To Limit Essential Viral Functions"

    Article Title: Human Adenovirus Core Protein V Is Targeted by the Host SUMOylation Machinery To Limit Essential Viral Functions

    Journal: Journal of Virology

    doi: 10.1128/JVI.01451-17

    Protein V is a novel SUMO target in the host cell. (A) HeLa cells and HeLa cells stably expressing 6His-SUMO1 or 6His-SUMO2 were transfected with 10 μg of an empty vector control or pCMX3b-Flag-V expression plasmid, harvested at 48 h p.t., and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates were purified using a Ni-NTA matrix (Ni-NTA pulldown), and input represents total cell lysates. Samples were separated by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using MAb M2 (anti-Flag), MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right (exp., exposure). (B) HeLa cells or HeLa cells stably expressing 6His-SUMO2 were transfected with 10 μg of an empty vector control or pCMX3b-Flag-V expression plasmid and superinfected with H5 pg 4100 (MOI of 20) at 24 h p.t. The cells were harvested at 24 h p.i. and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates purified by Ni-NTA pulldown, and input total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using pAb anti-V, MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.
    Figure Legend Snippet: Protein V is a novel SUMO target in the host cell. (A) HeLa cells and HeLa cells stably expressing 6His-SUMO1 or 6His-SUMO2 were transfected with 10 μg of an empty vector control or pCMX3b-Flag-V expression plasmid, harvested at 48 h p.t., and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates were purified using a Ni-NTA matrix (Ni-NTA pulldown), and input represents total cell lysates. Samples were separated by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using MAb M2 (anti-Flag), MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right (exp., exposure). (B) HeLa cells or HeLa cells stably expressing 6His-SUMO2 were transfected with 10 μg of an empty vector control or pCMX3b-Flag-V expression plasmid and superinfected with H5 pg 4100 (MOI of 20) at 24 h p.t. The cells were harvested at 24 h p.i. and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates purified by Ni-NTA pulldown, and input total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using pAb anti-V, MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.

    Techniques Used: Stable Transfection, Expressing, Transfection, Plasmid Preparation, Purification, SDS Page, Labeling

    Identification of protein V SUMOylation sites. (A) In silico analysis of protein V to determine potential SUMO conjugation or interaction motifs, using the algorithms SUMOPlot (Abgent, Inc., San Diego, CA), GPS-SUMO, and Jassa. (B) Schematic illustration of the protein V SCM mutants used in the experiments. SCMs are depicted in pink, nSCMs are in green, and K → R exchanges are in yellow. (C) HeLa cells or HeLa cells stably expressing 6His-SUMO2 were transfected with either 10 μg of an empty vector control, pCMX3b-Flag-V expression plasmid, or Flag-V SCM mutants as indicated. Cells were harvested at 48 h p.t. and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates purified by using Ni-NTA pulldown and input of total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using MAb M2 (anti-Flag), MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.
    Figure Legend Snippet: Identification of protein V SUMOylation sites. (A) In silico analysis of protein V to determine potential SUMO conjugation or interaction motifs, using the algorithms SUMOPlot (Abgent, Inc., San Diego, CA), GPS-SUMO, and Jassa. (B) Schematic illustration of the protein V SCM mutants used in the experiments. SCMs are depicted in pink, nSCMs are in green, and K → R exchanges are in yellow. (C) HeLa cells or HeLa cells stably expressing 6His-SUMO2 were transfected with either 10 μg of an empty vector control, pCMX3b-Flag-V expression plasmid, or Flag-V SCM mutants as indicated. Cells were harvested at 48 h p.t. and subjected to a guanidinium chloride buffer. 6His-SUMO conjugates purified by using Ni-NTA pulldown and input of total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Ni-NTA-purified and input proteins were detected using MAb M2 (anti-Flag), MAb anti-6His, and MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.

    Techniques Used: In Silico, Conjugation Assay, Stable Transfection, Expressing, Transfection, Plasmid Preparation, Purification, SDS Page, Labeling

    HAdV protein V association with host nucleoli structures. (A) HepaRG cells were infected with H5 pg 4100 (MOI of 20) and fixed with 4% PFA at 24 h p.i. Mock, uninfected control. Proteins were detected with pAb anti-protein V and MAb FC-61991 (anti-B23). Primary antibodies were detected with secondary antibodies conjugated with Alexa 488 (green) or Cy3 (red), and nuclei were stained with DAPI (blue). Merged images show the overlay of single images of each row. Images were captured with a Nikon confocal fluorescence microscope. Below these images, colocalization of adenoviral protein V and B23 at host nucleoli is depicted by two-dimensional histograms, which correlate the pixel intensities of two channels and show the corresponding channel overlay of the analyzed regions of interest. tM is the thresholded Mander's split coefficient, and the number indicates the channel, with 1 corresponding to the red channel (B23) and 2 corresponding to the green channel (protein V). (B) H1299 cells were transfected with either 10 μg of an empty vector control, a pCMX3b-Flag-V expression plasmid (left panel), or a pcDNA3-HA-V expression plasmid. Cells were harvested at 48 h p.t. Total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Levels of Flag-V and HA-V were detected by using MAb M2 (anti-Flag) and MAb 3F10 (anti-HA); input levels of β-actin were detected by using MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.
    Figure Legend Snippet: HAdV protein V association with host nucleoli structures. (A) HepaRG cells were infected with H5 pg 4100 (MOI of 20) and fixed with 4% PFA at 24 h p.i. Mock, uninfected control. Proteins were detected with pAb anti-protein V and MAb FC-61991 (anti-B23). Primary antibodies were detected with secondary antibodies conjugated with Alexa 488 (green) or Cy3 (red), and nuclei were stained with DAPI (blue). Merged images show the overlay of single images of each row. Images were captured with a Nikon confocal fluorescence microscope. Below these images, colocalization of adenoviral protein V and B23 at host nucleoli is depicted by two-dimensional histograms, which correlate the pixel intensities of two channels and show the corresponding channel overlay of the analyzed regions of interest. tM is the thresholded Mander's split coefficient, and the number indicates the channel, with 1 corresponding to the red channel (B23) and 2 corresponding to the green channel (protein V). (B) H1299 cells were transfected with either 10 μg of an empty vector control, a pCMX3b-Flag-V expression plasmid (left panel), or a pcDNA3-HA-V expression plasmid. Cells were harvested at 48 h p.t. Total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Levels of Flag-V and HA-V were detected by using MAb M2 (anti-Flag) and MAb 3F10 (anti-HA); input levels of β-actin were detected by using MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.

    Techniques Used: Infection, Staining, Fluorescence, Microscopy, Transfection, Plasmid Preparation, Expressing, SDS Page, Labeling

    5) Product Images from "Btf, a Novel Death-Promoting Transcriptional Repressor That Interacts with Bcl-2-Related Proteins"

    Article Title: Btf, a Novel Death-Promoting Transcriptional Repressor That Interacts with Bcl-2-Related Proteins

    Journal: Molecular and Cellular Biology

    doi:

    Nuclear subcellular localization of Btf S is altered by E1B 19K, Bcl-2, and Btf L . (A) HeLa cells were transfected with expression plasmid pcDNA3-Myc-Btf S alone or with pCMV E1B 19K or pcDNA3–Bcl-2 as indicated. The cells were fixed 24 h posttransfection and double stained against Myc and E1B 19K or Bcl-2. Magnification, ×1,000. (B) HeLa cells expressing Bcl-x L were transfected with pcDNA3-Myc-Btf S and stained against Myc 24 h posttransfection. Values represent the percentages of nuclear Btf S expression (Btf S /total Btf S ) in these cells.
    Figure Legend Snippet: Nuclear subcellular localization of Btf S is altered by E1B 19K, Bcl-2, and Btf L . (A) HeLa cells were transfected with expression plasmid pcDNA3-Myc-Btf S alone or with pCMV E1B 19K or pcDNA3–Bcl-2 as indicated. The cells were fixed 24 h posttransfection and double stained against Myc and E1B 19K or Bcl-2. Magnification, ×1,000. (B) HeLa cells expressing Bcl-x L were transfected with pcDNA3-Myc-Btf S and stained against Myc 24 h posttransfection. Values represent the percentages of nuclear Btf S expression (Btf S /total Btf S ) in these cells.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Staining

    BP-1 interacts with E1B 19K in yeast. (A) The yeast two-hybrid assay was used to demonstrate binding between BP-1 and E1B 19K. Growth in the presence of histidine indicates that both plasmids can be expressed in yeast, and growth in the absence of histidine demonstrates an interaction between the two proteins. Apc-2 represents an irrelevant hydrophobic protein used as a negative control. The specificity of the interaction between BP-1 and E1B 19K was tested by using related proteins (Bcl-2, Bcl-x L , and Bax) as well as missense mutants of E1B 19K (pm7, pm51, pm87, and pm102). (B) The minimal regions of E1B 19K required for interaction with BP-1 were mapped by using a series of deletion mutants. (C) Schematic representation of E1B 19K showing the missense and deletion mutants tested in the two-hybrid assay. Regions I and III indicate the locations of the Bcl-2 homology domains BH1 and BH3, respectively. E1B 19K does not contain recognizable BH2 or BH4 domains.
    Figure Legend Snippet: BP-1 interacts with E1B 19K in yeast. (A) The yeast two-hybrid assay was used to demonstrate binding between BP-1 and E1B 19K. Growth in the presence of histidine indicates that both plasmids can be expressed in yeast, and growth in the absence of histidine demonstrates an interaction between the two proteins. Apc-2 represents an irrelevant hydrophobic protein used as a negative control. The specificity of the interaction between BP-1 and E1B 19K was tested by using related proteins (Bcl-2, Bcl-x L , and Bax) as well as missense mutants of E1B 19K (pm7, pm51, pm87, and pm102). (B) The minimal regions of E1B 19K required for interaction with BP-1 were mapped by using a series of deletion mutants. (C) Schematic representation of E1B 19K showing the missense and deletion mutants tested in the two-hybrid assay. Regions I and III indicate the locations of the Bcl-2 homology domains BH1 and BH3, respectively. E1B 19K does not contain recognizable BH2 or BH4 domains.

    Techniques Used: Y2H Assay, Binding Assay, Negative Control, Two Hybrid Assay

    Reporter assay demonstrating that Btf S is a transcription repressor and is inhibited by Bcl-2-like proteins. HeLa cells were transfected with 2.5 μg of the reporter construct (luciferase construct containing GAL4 DNA binding sites within its promoter), 2.5 μg of GAL4 DNA binding domain fusion genes (pm1-Btf S , pm1-ΔN552, and pm1-ΔC210 or empty pm1 vector control), and 2.5 μg of bcl-2 family gene (pCMV-E1B 19K, pcDNA3–Bcl-2, pcDNA3-Flag–Bcl-x L , or empty pcDNA3 vector control). The cells were harvested 24 hours posttransfection, and the luciferase activity was measured in a scintillation counter with the luciferase substrate luciferin. Values were normalized for protein concentrations measured by the Bradford assay and graphed as a percentage of the result for the negative control (empty pm1 vector). The experiment was performed six times, and the high and low values for each sample were dropped. Bars indicate standard deviations ( n = 4).
    Figure Legend Snippet: Reporter assay demonstrating that Btf S is a transcription repressor and is inhibited by Bcl-2-like proteins. HeLa cells were transfected with 2.5 μg of the reporter construct (luciferase construct containing GAL4 DNA binding sites within its promoter), 2.5 μg of GAL4 DNA binding domain fusion genes (pm1-Btf S , pm1-ΔN552, and pm1-ΔC210 or empty pm1 vector control), and 2.5 μg of bcl-2 family gene (pCMV-E1B 19K, pcDNA3–Bcl-2, pcDNA3-Flag–Bcl-x L , or empty pcDNA3 vector control). The cells were harvested 24 hours posttransfection, and the luciferase activity was measured in a scintillation counter with the luciferase substrate luciferin. Values were normalized for protein concentrations measured by the Bradford assay and graphed as a percentage of the result for the negative control (empty pm1 vector). The experiment was performed six times, and the high and low values for each sample were dropped. Bars indicate standard deviations ( n = 4).

    Techniques Used: Reporter Assay, Transfection, Construct, Luciferase, Binding Assay, Plasmid Preparation, Activity Assay, Bradford Assay, Negative Control

    6) Product Images from "Human Adenovirus Core Protein V Is Targeted by the Host SUMOylation Machinery To Limit Essential Viral Functions"

    Article Title: Human Adenovirus Core Protein V Is Targeted by the Host SUMOylation Machinery To Limit Essential Viral Functions

    Journal: Journal of Virology

    doi: 10.1128/JVI.01451-17

    HAdV protein V association with host nucleoli structures. (A) HepaRG cells were infected with H5 pg 4100 (MOI of 20) and fixed with 4% PFA at 24 h p.i. Mock, uninfected control. Proteins were detected with pAb anti-protein V and MAb FC-61991 (anti-B23). Primary antibodies were detected with secondary antibodies conjugated with Alexa 488 (green) or Cy3 (red), and nuclei were stained with DAPI (blue). Merged images show the overlay of single images of each row. Images were captured with a Nikon confocal fluorescence microscope. Below these images, colocalization of adenoviral protein V and B23 at host nucleoli is depicted by two-dimensional histograms, which correlate the pixel intensities of two channels and show the corresponding channel overlay of the analyzed regions of interest. tM is the thresholded Mander's split coefficient, and the number indicates the channel, with 1 corresponding to the red channel (B23) and 2 corresponding to the green channel (protein V). (B) H1299 cells were transfected with either 10 μg of an empty vector control, a pCMX3b-Flag-V expression plasmid (left panel), or a pcDNA3-HA-V expression plasmid. Cells were harvested at 48 h p.t. Total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Levels of Flag-V and HA-V were detected by using MAb M2 (anti-Flag) and MAb 3F10 (anti-HA); input levels of β-actin were detected by using MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.
    Figure Legend Snippet: HAdV protein V association with host nucleoli structures. (A) HepaRG cells were infected with H5 pg 4100 (MOI of 20) and fixed with 4% PFA at 24 h p.i. Mock, uninfected control. Proteins were detected with pAb anti-protein V and MAb FC-61991 (anti-B23). Primary antibodies were detected with secondary antibodies conjugated with Alexa 488 (green) or Cy3 (red), and nuclei were stained with DAPI (blue). Merged images show the overlay of single images of each row. Images were captured with a Nikon confocal fluorescence microscope. Below these images, colocalization of adenoviral protein V and B23 at host nucleoli is depicted by two-dimensional histograms, which correlate the pixel intensities of two channels and show the corresponding channel overlay of the analyzed regions of interest. tM is the thresholded Mander's split coefficient, and the number indicates the channel, with 1 corresponding to the red channel (B23) and 2 corresponding to the green channel (protein V). (B) H1299 cells were transfected with either 10 μg of an empty vector control, a pCMX3b-Flag-V expression plasmid (left panel), or a pcDNA3-HA-V expression plasmid. Cells were harvested at 48 h p.t. Total cell lysates were resolved by SDS-PAGE and visualized by immunoblotting. Levels of Flag-V and HA-V were detected by using MAb M2 (anti-Flag) and MAb 3F10 (anti-HA); input levels of β-actin were detected by using MAb AC-15 (anti-β-actin). Molecular masses in kilodaltons are indicated on the left, while corresponding proteins are labeled on the right.

    Techniques Used: Infection, Staining, Fluorescence, Microscopy, Transfection, Plasmid Preparation, Expressing, SDS Page, Labeling

    7) Product Images from "Evaluation of apoptogenic adenovirus type 5 oncolytic vectors in a Syrian hamster head and neck cancer model"

    Article Title: Evaluation of apoptogenic adenovirus type 5 oncolytic vectors in a Syrian hamster head and neck cancer model

    Journal: Cancer gene therapy

    doi: 10.1038/cgt.2014.22

    HAdV5 mutants and replication in normal cells (A). Diagrammatic illustration of HAdV5 mutants. The top panel shows the proteins expressed from early regions E1B and E3. Mutants lp 11w and lp 11w/Δ55K express wt E1A and E3 regions while the control mutant dl 312 contain deletions in these regions. The mutations within the E1B region are indicated. (B). Expression of E1A and E1B proteins by HAdV5 mutants. (C). Replication of HAdV5 mutants in normal human epithelial (NHBE) and fibroblast (HFF) cells. The virus yield from infected NHBE and HFF cells were determined by plaque assay on 293 cells.
    Figure Legend Snippet: HAdV5 mutants and replication in normal cells (A). Diagrammatic illustration of HAdV5 mutants. The top panel shows the proteins expressed from early regions E1B and E3. Mutants lp 11w and lp 11w/Δ55K express wt E1A and E3 regions while the control mutant dl 312 contain deletions in these regions. The mutations within the E1B region are indicated. (B). Expression of E1A and E1B proteins by HAdV5 mutants. (C). Replication of HAdV5 mutants in normal human epithelial (NHBE) and fibroblast (HFF) cells. The virus yield from infected NHBE and HFF cells were determined by plaque assay on 293 cells.

    Techniques Used: Mutagenesis, Expressing, Infection, Plaque Assay

    8) Product Images from "Low-Level Expression of the E1B 20-Kilodalton Protein by Adenovirus 14p1 Enhances Viral Immunopathogenesis"

    Article Title: Low-Level Expression of the E1B 20-Kilodalton Protein by Adenovirus 14p1 Enhances Viral Immunopathogenesis

    Journal: Journal of Virology

    doi: 10.1128/JVI.01790-15

    Reduced E1B 20K expression correlated with loss of Ad14p1 CPE corpse repression of NF-κB activation. (A) Immunoblot analysis of E1B 19/20K expression in A549 cells infected with wt Ad14, wt Ad5, or Ad14p1 clinical isolate 1986T. (B) RT-qPCR analysis
    Figure Legend Snippet: Reduced E1B 20K expression correlated with loss of Ad14p1 CPE corpse repression of NF-κB activation. (A) Immunoblot analysis of E1B 19/20K expression in A549 cells infected with wt Ad14, wt Ad5, or Ad14p1 clinical isolate 1986T. (B) RT-qPCR analysis

    Techniques Used: Expressing, Activation Assay, Infection, Quantitative RT-PCR

    9) Product Images from "DNA damage response and MCL-1 destruction initiate apoptosis in adenovirus-infected cells"

    Article Title: DNA damage response and MCL-1 destruction initiate apoptosis in adenovirus-infected cells

    Journal: Genes & Development

    doi: 10.1101/gad.1156903

    E1A expression during adenovirus infection causes the loss of MCL-1 protein. ( A ) HeLa cells were mock, Ad5 dl 309, Ad5 dl 337, Ad5E1B - , Ad5E1A - , or Pac3 infected, and representative samples were collected at 0 (for mock only), 12, 24, 36, 48, and 72 h post-infection, and analyzed by Western blotting with antibodies against MCL-1, BCL-2, and actin. ( B ) Adenovirus infection stimulates proteasome-mediated turnover of MCL-1. HeLa cells were mock, Ad5 dl 309, or Ad5 dl 337 infected for 24 h, and then treated with cycloheximide (CHX) for 0, 2, 4, 6, and 20 h, or a combination of CHX plus epoxomicin (EPO) for 6 or 20 h, or were left untreated (UT). Samples were collected at the indicated time points and analyzed by Western blotting with anti-MCL-1 and anti-actin antibodies. ( C ) The effect of adenovirus infection on mcl-1 mRNA levels. Total RNA from mock, Ad5 dl 309, Ad5 dl 337, or Ad5E1A - infected cells was isolated at 0, 14, and 24 h post-infection, and was analyzed by Taqman real-time PCR using a probe and primers specific for mcl-1 mRNA. Fold change represents the difference in fluorescence signal in the 14- and 24-h samples compared with the corresponding 0-h sample. Error bars, the standard deviation of triplicate samples. Samples were also analyzed by Taqman real-time PCR with primers and a probe specific for gapdh .
    Figure Legend Snippet: E1A expression during adenovirus infection causes the loss of MCL-1 protein. ( A ) HeLa cells were mock, Ad5 dl 309, Ad5 dl 337, Ad5E1B - , Ad5E1A - , or Pac3 infected, and representative samples were collected at 0 (for mock only), 12, 24, 36, 48, and 72 h post-infection, and analyzed by Western blotting with antibodies against MCL-1, BCL-2, and actin. ( B ) Adenovirus infection stimulates proteasome-mediated turnover of MCL-1. HeLa cells were mock, Ad5 dl 309, or Ad5 dl 337 infected for 24 h, and then treated with cycloheximide (CHX) for 0, 2, 4, 6, and 20 h, or a combination of CHX plus epoxomicin (EPO) for 6 or 20 h, or were left untreated (UT). Samples were collected at the indicated time points and analyzed by Western blotting with anti-MCL-1 and anti-actin antibodies. ( C ) The effect of adenovirus infection on mcl-1 mRNA levels. Total RNA from mock, Ad5 dl 309, Ad5 dl 337, or Ad5E1A - infected cells was isolated at 0, 14, and 24 h post-infection, and was analyzed by Taqman real-time PCR using a probe and primers specific for mcl-1 mRNA. Fold change represents the difference in fluorescence signal in the 14- and 24-h samples compared with the corresponding 0-h sample. Error bars, the standard deviation of triplicate samples. Samples were also analyzed by Taqman real-time PCR with primers and a probe specific for gapdh .

    Techniques Used: Expressing, Infection, Western Blot, Isolation, Real-time Polymerase Chain Reaction, Fluorescence, Standard Deviation

    Related Articles

    Western Blot:

    Article Title: Chromatin remodeller Fun30Fft3 induces nucleosome disassembly to facilitate RNA polymerase II elongation
    Article Snippet: .. Antibodies Antibodies against α-RNA polymerase II CTD repeat YSPTSPS without phosphorylation (8WG16; 920102, BioLegend; previously MMS-126R, Covance; 0.1 μg μl−1 ), the α-RNA polymerase II CTD repeat YSPTSPS with phosphorylation at Ser2 (phospho S2; ab5095, Abcam; 1 μg μl−1 ), α-FLAG M2 (F1804, Sigma; 1 μg μl−1 ), and α-beta actin (sc-47778, Santa Cruz; 0.2 μg μl−1 ) were used for ChIP-seq, co-immunoprecipitation and western blot. .. In addition, the α-H3 antibody (rabbit polyclonal; serum; 1 μg μl−1 ), which was produced in-house against recombinant yeast histone H3 , were also used for ChIP-seq and western blot.

    Article Title: CaMKII-mediated Beclin 1 phosphorylation regulates autophagy that promotes degradation of Id and neuroblastoma cell differentiation
    Article Snippet: .. Other primary antibodies used for western blotting were anti-Beclin 1 (Cell Signaling Technology, #3738, 1:1000), anti-GAPDH (KangChen, Shanghai, China, 1:10,000), anti-p-CaMKII (Cell Signaling Technology, #3361, 1:1000), anti-CaMKII (Santa Cruz, sc-1541, 1:500), anti-Bcl-2 (Santa Cruz, SC-7382, 1:500), anti-LC3 (Novus, Littleton, CO, USA, NB100-2220, 1:3000), anti-Id-1 (Santa Cruz, sc-488, 1:1000), anti-Id-2 (Cell Signaling Technology, #3431, 1:500), anti-SQSTM1 (Santa Cruz, sc-28359, 1:1000), anti-His-Tag (Cell Signaling Technology, #2366, 1:1000), anti-Myc-Tag (Cell Signaling Technology, #2276, 1:1000), anti-Flag (Sigma-Aldrich, F1804, 1:2000), anti-ubiquitin (Santa Cruz, sc-58449, 1:1000), anti-K63-linkage-specific polyubiquitin (Cell Signaling Technology, #5621, 1:1000), anti-TRAF-6 (Cell Signaling Technology, #8028, 1:1000), anti-GAP43 (Cell Signaling Technology, #8945 s, 1:1000), anti-NF68 (Cell Signaling Technology, #2837 s, 1:1000), anti-nestin (Santa Cruz, SC-23927, 1:1000), anti-vimentin (BD, 550513, 1:1000), and anti-E-cadherin (BD, 51-9001922, 1:1000). .. KN-93, MG132, and bafilomycin A1 were purchased from Sigma-Aldrich.

    Immunohistochemistry:

    Article Title: Methylation-dependent regulation of HIF-1α stability restricts retinal and tumour angiogenesis
    Article Snippet: .. Antibodies The following commercially available antibodies were used: anti-HIF-1α (NB100–132, Novus; 10006421, 1:1,000 dilution for IB analysis, Cayman; 1:1,000 dilution for IB analysis, 1:200 for IF analysis; MAB 1536, R & D Systems, 1:1,000 dilution for IB analysis); anti-HIF-2α (NB100–122, Novus, 1:1,000 dilution for IB anlysis); anti-Xpress (R910-25, Invitrogen, 1:5,000 dilution for IB analysis); anti-FLAG (F3165, Sigma, 1:10,000 dilution for IB analysis); anti-methyl-Lys (ab23366, Abcam); anti-CD31 (clone 2H8, MAB1398Z, Millipore, 1:200 dilution for immunohistochemical (IHC) analysis); anti-HA (MMS-101R, Covance, 1:5,000 dilution for IB analysis); anti-VEGF (AF493NA, R & D System, 1:200 dilution for IHC analysis); anti-EPO (sc-7956, 1:1,000 for IB analysis), anti-Brn3b (sc-6026, 1:200 dilution for IHC analysis) from Santa Cruz; anti-LSD1 (#2139, 1:1,000 dilution for IB analysis), anti-hydroxyl-HIF-1α (#3434, 1:5,000 dilution for IB analysis), anti-Caspase3 (#9661, 1:200 dilution for IHC analysis), anti-Ki-67 (#9027, 1:100 dilution for IHC analysis) and anti-SET7/9 antibodies (#2813, 1:1,000 dilution for IB analysis) from Cell Signalling. .. Anti-HIF-1α-K32 methyl antibodies were generated by Abfrontier (South Korea, 1:5,000 dilution for IB analysis).

    other:

    Article Title: NMDA receptors are selectively partitioned into complexes and supercomplexes during synapse maturation
    Article Snippet: Antibodies Primary antibodies (dilutions for immunoblot in parenthesis: mouse Flag (Sigma, F3165) mouse GluN1 (1:1,000, Invitrogen, 32–0500), rabbit GluN1-1 (1:500, Millipore, AB9864), mouse GluN2A (1:1,000, BD Bioscience, 612287), rabbit GluN2A (1:500, Serotec, AHP1880), mouse GluN2B (1:1,000, BD Bioscience, 610416/7), rabbit GluN2B (1:500, Invitrogen, 71–8,600), mouse PSD95/3 (1:3,000, Thermo scientific, MA1–045), mouse PSD93 (1:2,500, Neuromab, 75-057), rabbit PSD95 (1:1,000.

    Article Title: Inverse Agonist and Pharmacochaperone Properties of MK-0524 on the Prostanoid DP1 Receptor
    Article Snippet: Reagents Monoclonal anti-FLAG (M2) (cat. F3165), monoclonal anti-FLAG (M1) (cat. F3040), and goat alkaline phosphatase-conjugated anti-mouse IgG (cat. A3562) antibodies were from Sigma-Aldrich, MO.

    ChIP-sequencing:

    Article Title: Chromatin remodeller Fun30Fft3 induces nucleosome disassembly to facilitate RNA polymerase II elongation
    Article Snippet: .. Antibodies Antibodies against α-RNA polymerase II CTD repeat YSPTSPS without phosphorylation (8WG16; 920102, BioLegend; previously MMS-126R, Covance; 0.1 μg μl−1 ), the α-RNA polymerase II CTD repeat YSPTSPS with phosphorylation at Ser2 (phospho S2; ab5095, Abcam; 1 μg μl−1 ), α-FLAG M2 (F1804, Sigma; 1 μg μl−1 ), and α-beta actin (sc-47778, Santa Cruz; 0.2 μg μl−1 ) were used for ChIP-seq, co-immunoprecipitation and western blot. .. In addition, the α-H3 antibody (rabbit polyclonal; serum; 1 μg μl−1 ), which was produced in-house against recombinant yeast histone H3 , were also used for ChIP-seq and western blot.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Millipore rabbit polyclonal anti e1b 19k
    Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or <t>E1B</t> <t>19K.</t> Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).
    Rabbit Polyclonal Anti E1b 19k, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti e1b 19k/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti e1b 19k - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    80
    Millipore e1b 19k production
    Nuclear subcellular localization of Btf S is altered by <t>E1B</t> <t>19K,</t> Bcl-2, and Btf L . (A) HeLa cells were transfected with expression plasmid pcDNA3-Myc-Btf S alone or with pCMV E1B 19K or pcDNA3–Bcl-2 as indicated. The cells were fixed 24 h posttransfection and double stained against Myc and E1B 19K or Bcl-2. Magnification, ×1,000. (B) HeLa cells expressing Bcl-x L were transfected with pcDNA3-Myc-Btf S and stained against Myc 24 h posttransfection. Values represent the percentages of nuclear Btf S expression (Btf S /total Btf S ) in these cells.
    E1b 19k Production, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e1b 19k production/product/Millipore
    Average 80 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    e1b 19k production - by Bioz Stars, 2020-09
    80/100 stars
      Buy from Supplier

    90
    Millipore ad14 e1b 20k
    Reduced <t>E1B</t> <t>20K</t> expression correlated with loss of Ad14p1 CPE corpse repression of NF-κB activation. (A) Immunoblot analysis of E1B 19/20K expression in A549 cells infected with wt <t>Ad14,</t> wt Ad5, or Ad14p1 clinical isolate 1986T. (B) RT-qPCR analysis
    Ad14 E1b 20k, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ad14 e1b 20k/product/Millipore
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ad14 e1b 20k - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).

    Journal: Genes & Development

    Article Title: Hypoxia and defective apoptosis drive genomic instability and tumorigenesis

    doi: 10.1101/gad.1204904

    Figure Lengend Snippet: Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).

    Article Snippet: Primary antibodies used were hamster anti-human BCL-2 (BD-Pharmingen); rabbit polyclonal anti-E1B 19K , mouse anti-actin (Oncogene Research Products), rabbit anti-mouse HIF-1α (Cayman), rabbit anti-BIM (Axxora), or rabbit polyclonal antisera raised in our lab against a GST-human PUMA fusion protein encoding a 102-amino acid region common to PUMA-α and PUMA-β.

    Techniques: Staining, Transformation Assay, Expressing, Injection, Derivative Assay, Immunohistochemistry, Mouse Assay

    Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.

    Journal: Genes & Development

    Article Title: Hypoxia and defective apoptosis drive genomic instability and tumorigenesis

    doi: 10.1101/gad.1204904

    Figure Lengend Snippet: Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.

    Article Snippet: Primary antibodies used were hamster anti-human BCL-2 (BD-Pharmingen); rabbit polyclonal anti-E1B 19K , mouse anti-actin (Oncogene Research Products), rabbit anti-mouse HIF-1α (Cayman), rabbit anti-BIM (Axxora), or rabbit polyclonal antisera raised in our lab against a GST-human PUMA fusion protein encoding a 102-amino acid region common to PUMA-α and PUMA-β.

    Techniques: Blocking Assay, Stable Transfection, Western Blot, Expressing, Clone Assay, Plasmid Preparation, Injection, Mouse Assay

    Nuclear subcellular localization of Btf S is altered by E1B 19K, Bcl-2, and Btf L . (A) HeLa cells were transfected with expression plasmid pcDNA3-Myc-Btf S alone or with pCMV E1B 19K or pcDNA3–Bcl-2 as indicated. The cells were fixed 24 h posttransfection and double stained against Myc and E1B 19K or Bcl-2. Magnification, ×1,000. (B) HeLa cells expressing Bcl-x L were transfected with pcDNA3-Myc-Btf S and stained against Myc 24 h posttransfection. Values represent the percentages of nuclear Btf S expression (Btf S /total Btf S ) in these cells.

    Journal: Molecular and Cellular Biology

    Article Title: Btf, a Novel Death-Promoting Transcriptional Repressor That Interacts with Bcl-2-Related Proteins

    doi:

    Figure Lengend Snippet: Nuclear subcellular localization of Btf S is altered by E1B 19K, Bcl-2, and Btf L . (A) HeLa cells were transfected with expression plasmid pcDNA3-Myc-Btf S alone or with pCMV E1B 19K or pcDNA3–Bcl-2 as indicated. The cells were fixed 24 h posttransfection and double stained against Myc and E1B 19K or Bcl-2. Magnification, ×1,000. (B) HeLa cells expressing Bcl-x L were transfected with pcDNA3-Myc-Btf S and stained against Myc 24 h posttransfection. Values represent the percentages of nuclear Btf S expression (Btf S /total Btf S ) in these cells.

    Article Snippet: The relative levels of BtfS and E1B 19K production were determined by immunoprecipitation with anti-Myc (Oncogene Research Products, Cambridge, Mass.) or anti-E1B 19K ( ) in NETN buffer followed by protein A-Sepharose and then washing in NETN.

    Techniques: Transfection, Expressing, Plasmid Preparation, Staining

    BP-1 interacts with E1B 19K in yeast. (A) The yeast two-hybrid assay was used to demonstrate binding between BP-1 and E1B 19K. Growth in the presence of histidine indicates that both plasmids can be expressed in yeast, and growth in the absence of histidine demonstrates an interaction between the two proteins. Apc-2 represents an irrelevant hydrophobic protein used as a negative control. The specificity of the interaction between BP-1 and E1B 19K was tested by using related proteins (Bcl-2, Bcl-x L , and Bax) as well as missense mutants of E1B 19K (pm7, pm51, pm87, and pm102). (B) The minimal regions of E1B 19K required for interaction with BP-1 were mapped by using a series of deletion mutants. (C) Schematic representation of E1B 19K showing the missense and deletion mutants tested in the two-hybrid assay. Regions I and III indicate the locations of the Bcl-2 homology domains BH1 and BH3, respectively. E1B 19K does not contain recognizable BH2 or BH4 domains.

    Journal: Molecular and Cellular Biology

    Article Title: Btf, a Novel Death-Promoting Transcriptional Repressor That Interacts with Bcl-2-Related Proteins

    doi:

    Figure Lengend Snippet: BP-1 interacts with E1B 19K in yeast. (A) The yeast two-hybrid assay was used to demonstrate binding between BP-1 and E1B 19K. Growth in the presence of histidine indicates that both plasmids can be expressed in yeast, and growth in the absence of histidine demonstrates an interaction between the two proteins. Apc-2 represents an irrelevant hydrophobic protein used as a negative control. The specificity of the interaction between BP-1 and E1B 19K was tested by using related proteins (Bcl-2, Bcl-x L , and Bax) as well as missense mutants of E1B 19K (pm7, pm51, pm87, and pm102). (B) The minimal regions of E1B 19K required for interaction with BP-1 were mapped by using a series of deletion mutants. (C) Schematic representation of E1B 19K showing the missense and deletion mutants tested in the two-hybrid assay. Regions I and III indicate the locations of the Bcl-2 homology domains BH1 and BH3, respectively. E1B 19K does not contain recognizable BH2 or BH4 domains.

    Article Snippet: The relative levels of BtfS and E1B 19K production were determined by immunoprecipitation with anti-Myc (Oncogene Research Products, Cambridge, Mass.) or anti-E1B 19K ( ) in NETN buffer followed by protein A-Sepharose and then washing in NETN.

    Techniques: Y2H Assay, Binding Assay, Negative Control, Two Hybrid Assay

    Reporter assay demonstrating that Btf S is a transcription repressor and is inhibited by Bcl-2-like proteins. HeLa cells were transfected with 2.5 μg of the reporter construct (luciferase construct containing GAL4 DNA binding sites within its promoter), 2.5 μg of GAL4 DNA binding domain fusion genes (pm1-Btf S , pm1-ΔN552, and pm1-ΔC210 or empty pm1 vector control), and 2.5 μg of bcl-2 family gene (pCMV-E1B 19K, pcDNA3–Bcl-2, pcDNA3-Flag–Bcl-x L , or empty pcDNA3 vector control). The cells were harvested 24 hours posttransfection, and the luciferase activity was measured in a scintillation counter with the luciferase substrate luciferin. Values were normalized for protein concentrations measured by the Bradford assay and graphed as a percentage of the result for the negative control (empty pm1 vector). The experiment was performed six times, and the high and low values for each sample were dropped. Bars indicate standard deviations ( n = 4).

    Journal: Molecular and Cellular Biology

    Article Title: Btf, a Novel Death-Promoting Transcriptional Repressor That Interacts with Bcl-2-Related Proteins

    doi:

    Figure Lengend Snippet: Reporter assay demonstrating that Btf S is a transcription repressor and is inhibited by Bcl-2-like proteins. HeLa cells were transfected with 2.5 μg of the reporter construct (luciferase construct containing GAL4 DNA binding sites within its promoter), 2.5 μg of GAL4 DNA binding domain fusion genes (pm1-Btf S , pm1-ΔN552, and pm1-ΔC210 or empty pm1 vector control), and 2.5 μg of bcl-2 family gene (pCMV-E1B 19K, pcDNA3–Bcl-2, pcDNA3-Flag–Bcl-x L , or empty pcDNA3 vector control). The cells were harvested 24 hours posttransfection, and the luciferase activity was measured in a scintillation counter with the luciferase substrate luciferin. Values were normalized for protein concentrations measured by the Bradford assay and graphed as a percentage of the result for the negative control (empty pm1 vector). The experiment was performed six times, and the high and low values for each sample were dropped. Bars indicate standard deviations ( n = 4).

    Article Snippet: The relative levels of BtfS and E1B 19K production were determined by immunoprecipitation with anti-Myc (Oncogene Research Products, Cambridge, Mass.) or anti-E1B 19K ( ) in NETN buffer followed by protein A-Sepharose and then washing in NETN.

    Techniques: Reporter Assay, Transfection, Construct, Luciferase, Binding Assay, Plasmid Preparation, Activity Assay, Bradford Assay, Negative Control

    Reduced E1B 20K expression correlated with loss of Ad14p1 CPE corpse repression of NF-κB activation. (A) Immunoblot analysis of E1B 19/20K expression in A549 cells infected with wt Ad14, wt Ad5, or Ad14p1 clinical isolate 1986T. (B) RT-qPCR analysis

    Journal: Journal of Virology

    Article Title: Low-Level Expression of the E1B 20-Kilodalton Protein by Adenovirus 14p1 Enhances Viral Immunopathogenesis

    doi: 10.1128/JVI.01790-15

    Figure Lengend Snippet: Reduced E1B 20K expression correlated with loss of Ad14p1 CPE corpse repression of NF-κB activation. (A) Immunoblot analysis of E1B 19/20K expression in A549 cells infected with wt Ad14, wt Ad5, or Ad14p1 clinical isolate 1986T. (B) RT-qPCR analysis

    Article Snippet: Actin (Sigma-Aldrich antibody) and Ad14 E1B 20K (mouse anti-Ad2 E1B 19K monoclonal antibody, 3D11; Calbiochem) were detected by immunoblotting using enhanced chemiluminescence (ECL).

    Techniques: Expressing, Activation Assay, Infection, Quantitative RT-PCR