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Santa Cruz Biotechnology anti his
In vitro sumoylation of TIF-1γ with E4-ORF3. <t>GST–TIF-1γΔC</t> (100 nM) was incubated with 50 nM E1, 250 nM E2, 50 μM <t>His</t> 6 -SUMO3, and the indicated concentrations of His 6 –E4-ORF3-WT or His 6 –E4-ORF3-L103A proteins at 37 °C for 90 min. Reaction mixtures were analyzed by Western blot with anti–TIF-1γ, anti-SUMO2/3, and anti–E4-ORF3 antibodies.
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Images

1) Product Images from "The adenovirus E4-ORF3 protein functions as a SUMO E3 ligase for TIF-1γ sumoylation and poly-SUMO chain elongation"

Article Title: The adenovirus E4-ORF3 protein functions as a SUMO E3 ligase for TIF-1γ sumoylation and poly-SUMO chain elongation

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1603872113

In vitro sumoylation of TIF-1γ with E4-ORF3. GST–TIF-1γΔC (100 nM) was incubated with 50 nM E1, 250 nM E2, 50 μM His 6 -SUMO3, and the indicated concentrations of His 6 –E4-ORF3-WT or His 6 –E4-ORF3-L103A proteins at 37 °C for 90 min. Reaction mixtures were analyzed by Western blot with anti–TIF-1γ, anti-SUMO2/3, and anti–E4-ORF3 antibodies.
Figure Legend Snippet: In vitro sumoylation of TIF-1γ with E4-ORF3. GST–TIF-1γΔC (100 nM) was incubated with 50 nM E1, 250 nM E2, 50 μM His 6 -SUMO3, and the indicated concentrations of His 6 –E4-ORF3-WT or His 6 –E4-ORF3-L103A proteins at 37 °C for 90 min. Reaction mixtures were analyzed by Western blot with anti–TIF-1γ, anti-SUMO2/3, and anti–E4-ORF3 antibodies.

Techniques Used: In Vitro, Incubation, Western Blot

In vitro sumoylation of TIF-1γ with E4-ORF3. ( A ) GST-tagged recombinant TIF-1γ protein (100 nM) was incubated with 50 nM E1, 250 nM E2, 50 μM His 6 -SUMO3, and the indicated concentrations of His 6 -E4-ORF3-WT or His 6 -E4-ORF3-L103A proteins at 37 °C for 60 min. Reaction mixtures were analyzed by Western blot with anti–TIF-1γ, anti-SUMO2/3, and anti–E4-ORF3 antibodies. ( B ) GST–TIF-1γΔC (100 nM) was used as a substrate and sumoylation was analyzed as described in A . ( C ) GST–TIF-1γΔC was incubated with (+WT and +L103A) or without (−) His 6 –E4-ORF3 for the indicated time periods. ( D ) GST–TIF-1γΔC was sumoylated at 37 °C for 60 min and incubated with 10 nM SENP1 catalytic domain at 20 °C for 10 min. A total of 3 μM His 6 –E4-ORF3 was used in C and D . ( E ) Schematic representation of TIF-1γ and TIF-1γΔC. RBCC contains the RING finger, B box, and coiled-coil domain; PB contains the plant homeodomain (PHD) and bromodomain.
Figure Legend Snippet: In vitro sumoylation of TIF-1γ with E4-ORF3. ( A ) GST-tagged recombinant TIF-1γ protein (100 nM) was incubated with 50 nM E1, 250 nM E2, 50 μM His 6 -SUMO3, and the indicated concentrations of His 6 -E4-ORF3-WT or His 6 -E4-ORF3-L103A proteins at 37 °C for 60 min. Reaction mixtures were analyzed by Western blot with anti–TIF-1γ, anti-SUMO2/3, and anti–E4-ORF3 antibodies. ( B ) GST–TIF-1γΔC (100 nM) was used as a substrate and sumoylation was analyzed as described in A . ( C ) GST–TIF-1γΔC was incubated with (+WT and +L103A) or without (−) His 6 –E4-ORF3 for the indicated time periods. ( D ) GST–TIF-1γΔC was sumoylated at 37 °C for 60 min and incubated with 10 nM SENP1 catalytic domain at 20 °C for 10 min. A total of 3 μM His 6 –E4-ORF3 was used in C and D . ( E ) Schematic representation of TIF-1γ and TIF-1γΔC. RBCC contains the RING finger, B box, and coiled-coil domain; PB contains the plant homeodomain (PHD) and bromodomain.

Techniques Used: In Vitro, Recombinant, Incubation, Western Blot

E4-ORF3 enhances TIF-1γ sumoylation and proteasomal degradation. ( A ) His 6 -tagged SUMO3-expressing HeLa cells were infected with increasing amounts of wild-type Ad5 [Ad5-WT, 0, 50, 100, 200, and 400 virus paticles per cell (P/cell)] and SUMO-conjugated proteins were analyzed at 8 hours post-infection (hpi) (pull-down). RanGAP1 was used as a control for SUMO capture. Total cell lysates were analyzed by Western blot (lysate). ( B ) Recombinant empty Ad-CMV (lanes 1 and 2, 900 P/cell) or Ad-CMV-HA-E4-ORF3 expression vector (lanes 3–5, 100, 300, and 900 P/cell) were used to infect His 6 -SUMO3-HeLa cells. At 10 hpi, SUMO conjugates were analyzed as in A . ( C ) HeLa and A549 cells were infected with empty Ad-CMV (1,000 P/cell) or Ad-CMV-HA-E4-ORF3 (200 and 1,000 P/cell) and treated with 20 μM MG132 at 1 hpi. At 10 hpi (HeLa) or 12 hpi (A549), cells were harvested and protein levels were determined by Western blot.
Figure Legend Snippet: E4-ORF3 enhances TIF-1γ sumoylation and proteasomal degradation. ( A ) His 6 -tagged SUMO3-expressing HeLa cells were infected with increasing amounts of wild-type Ad5 [Ad5-WT, 0, 50, 100, 200, and 400 virus paticles per cell (P/cell)] and SUMO-conjugated proteins were analyzed at 8 hours post-infection (hpi) (pull-down). RanGAP1 was used as a control for SUMO capture. Total cell lysates were analyzed by Western blot (lysate). ( B ) Recombinant empty Ad-CMV (lanes 1 and 2, 900 P/cell) or Ad-CMV-HA-E4-ORF3 expression vector (lanes 3–5, 100, 300, and 900 P/cell) were used to infect His 6 -SUMO3-HeLa cells. At 10 hpi, SUMO conjugates were analyzed as in A . ( C ) HeLa and A549 cells were infected with empty Ad-CMV (1,000 P/cell) or Ad-CMV-HA-E4-ORF3 (200 and 1,000 P/cell) and treated with 20 μM MG132 at 1 hpi. At 10 hpi (HeLa) or 12 hpi (A549), cells were harvested and protein levels were determined by Western blot.

Techniques Used: Expressing, Infection, Western Blot, Recombinant, Plasmid Preparation

2) Product Images from "Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression"

Article Title: Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.24.13.5639-5649.2004

Menin interacts with wild-type and oncogenic MLL proteins. (A and B) 293 cells were transiently transfected with expression vectors encoding various MLL deletion or fusion mutants (shown schematically in panel A) containing HIS and FLAG epitope tags at their N termini. Nuclear extracts prepared from transfectants were subjected to immunoprecipitation (IP) with anti-FLAG antibody (M2). Immune precipitates were analyzed by SDS-PAGE and were immunoblotted with antibodies indicated to the right of the respective panels (for panel B, anti-His [D-8] for various MLL mutants, anti-MLL C [mmC2.1], anti-HCF-1 C [H12], and antimenin [C19], respectively). Positions of molecular size markers are indicated on the left. Coprecipitation of endogenous menin with exogenous MLL (+ or −) is indicated to the right of the schematic. (C) Nuclear extracts of REH and HB cells were subjected to immunoprecipitation using antimenin antibody (C19) or a negative control antibody (anti-DmMyb). Immune precipitates were analyzed by SDS-PAGE and were immunoblotted with anti-MLL N (mmN4.4, top gel), anti-MLL C (mmC2.1, middle gel), and anti-ENL antibodies (bottom gel) as indicated to the right of the panels. Wild-type MLL coprecipitated with menin in REH cells (lane 3). Both wild-type and fusion (MLL-ENL) MLL proteins coprecipitated with menin in HB cells (lane 6).
Figure Legend Snippet: Menin interacts with wild-type and oncogenic MLL proteins. (A and B) 293 cells were transiently transfected with expression vectors encoding various MLL deletion or fusion mutants (shown schematically in panel A) containing HIS and FLAG epitope tags at their N termini. Nuclear extracts prepared from transfectants were subjected to immunoprecipitation (IP) with anti-FLAG antibody (M2). Immune precipitates were analyzed by SDS-PAGE and were immunoblotted with antibodies indicated to the right of the respective panels (for panel B, anti-His [D-8] for various MLL mutants, anti-MLL C [mmC2.1], anti-HCF-1 C [H12], and antimenin [C19], respectively). Positions of molecular size markers are indicated on the left. Coprecipitation of endogenous menin with exogenous MLL (+ or −) is indicated to the right of the schematic. (C) Nuclear extracts of REH and HB cells were subjected to immunoprecipitation using antimenin antibody (C19) or a negative control antibody (anti-DmMyb). Immune precipitates were analyzed by SDS-PAGE and were immunoblotted with anti-MLL N (mmN4.4, top gel), anti-MLL C (mmC2.1, middle gel), and anti-ENL antibodies (bottom gel) as indicated to the right of the panels. Wild-type MLL coprecipitated with menin in REH cells (lane 3). Both wild-type and fusion (MLL-ENL) MLL proteins coprecipitated with menin in HB cells (lane 6).

Techniques Used: Transfection, Expressing, FLAG-tag, Immunoprecipitation, SDS Page, Negative Control

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Clone Assay:

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Centrifugation:

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Article Snippet: The beads were collected by centrifugation and then washed five times with buffer containing 25 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM DTT, and 1% Triton X-100. .. Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

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Amplification:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
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Blocking Assay:

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Article Snippet: .. Membranes were incubated with blocking buffer (Tris-buffered saline Tween 20 (TBST) with 5% skim milk) for 1 h and then probed with anti-His or anti-GST antibody (1:1000) (Santa Cruz biotechnology) diluted in blocking buffer overnight on a rotating platform at 4 °C. .. Blots were subsequently washed 3 times with TBST and then incubated with horseradish peroxidase-conjugated secondary antibody (1:2000) (Promega) diluted in TBST for 1 h at room temperature.

Incubation:

Article Title: Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression
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Article Title: The Proline-rich N-terminal Domain of G18 Exhibits a Novel G Protein Regulatory Function *
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Article Snippet: 2× SDS sample buffer (25 µL) was added to the washed beads, and the microfuge tubes were incubated in boiling water for 5 min. .. Anti-His (catalog no. SC-803; Santa Cruz Biotechnology), anti-Myc (catalog no. SC-40; Santa Cruz Biotechnology), and anti- (catalog no. A190-108A; Bethyl Laboratories) antibodies were used for immunoprecipitation and immunoblotting analyses according to the manufacturers’ protocols.

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Luciferase:

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In Silico:

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Expressing:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: Briefly, 5×106 HCT116/MMTV cells grown in 15 cm-dishes were transfected with 1 μg/well of plasmid expressing the His-tagged full-length, Δ181–225, 181–225 fragment of Set/TAF-Iβ or the His-tagged Set-Can, together with 10 μg/well of pRShGRα. .. They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.).

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Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. The His-tagged expression vectors of full-length and domain-truncated mutants of hCaMKIINα (as illustrated in ), including pKIINα, pKIINα1–41 , pKIINα1–53 , pKIINα1–68 , and the FLAG-tagged expression vectors of CaMKIIα (pFLAG-CaMKII), and CaMKIIα with H282 mutated to R (H282R) were constructed by PCR cloning and PCR mutation.

Modification:

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Article Snippet: HeLa cell lines were cultivated in 10% fetal calf serum (FCS)-supplemented Dulbecco’s modified Eagle’s medium (DMEM). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
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Western Blot:

Article Title: Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression
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Article Snippet: .. Western blotting and immunofluorescence were performed using the following primary antibodies: anti-FLAG (M2; F3165; Sigma), anti-His (H15; sc-803; Santa Cruz Biotechnology), anti-glutathione S -transferase (anti-GST; B-14; sc-138; Santa Cruz Biotechnology), anti-Gα12 (sc-409; Santa Cruz Biotechnology), anti-Myc (9E10; sc-40; Santa Cruz Biotechnology), anti-GM130 (610822; Becton Dickinson), anti-Myc (610990; Cell Signaling), and antihemagglutinin (anti-HA; H6908; Sigma). .. Horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit secondary antibodies were from Amersham Pharmacia Biotech.

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
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Article Snippet: Paragraph title: Western blot analysis. ... The commercial antibodies anti-DnaK (MBL International), horseradish peroxidase (HRP)-conjugated anti-HA (Roche), anti-MBP (New England BioLabs), anti-His (Santa Cruz Biotechnology), and anti-FLAG (Sigma) were used according to the manufacturers' instructions.

Transformation Assay:

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: The fusion constructs were transformed into Escherichia coli strain BL21. .. Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

Electron Microscopy:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: Paragraph title: Immunofluorescence and electron microscopy analysis ... The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Transfection:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: Transient transfections of these cells were performed using Effectene (Qiagen). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: Briefly, 5×106 HCT116/MMTV cells grown in 15 cm-dishes were transfected with 1 μg/well of plasmid expressing the His-tagged full-length, Δ181–225, 181–225 fragment of Set/TAF-Iβ or the His-tagged Set-Can, together with 10 μg/well of pRShGRα. .. They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.).

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: For AMPK-dependent in vivo phosphorylation of Med1, HEK293 cells, HeLa cells, and mouse primary hepatocytes were infected with Ad-Med1 (3.5 × 106 virus particle) or transiently transfected with 24 μg of pShuttle-His-Med1 plasmid using Lipofectamine (Invitrogen). .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Chromatography:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. An hCaMKIINα-specific polyclonal antibody was raised against affinity chromatography-purified GST-KIINα1–41 fusion protein, and the antibody specificity was confirmed by Western blot and immunohistochemical staining for no cross-reactivity with hCaMKIINβ.

Concentration Assay:

Article Title: Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression
Article Snippet: Nuclear extract (5 ml) was incubated with primary antibodies at a concentration of approximately 1 μg/200 μl of extract or 50 μl of half the slurry of FLAG M2-agarose affinity gel and was used for immunoprecipitations and Western blotting as described elsewhere ( ). .. Anti-menin (H-300 or C19), anti-mSin3A (K-20), anti-SNF2H (H-300), anti-HIS (D-8), and anti-BRM (N-19) antibodies were purchased from Santa Cruz Biotechnology, Inc. Anti-RBBP5 (BL766) was purchased from Bethyl Laboratories, Inc. Anti-Flag (M2) agarose affinity gel was purchased from Sigma.

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: Transformants were grown to a concentration of OD600 = 0.6 in the 37°C shaker and then induced to express the fusion protein by incubation in growth culture supplemented with 0.5 mM isopropyl β- d -1-thiogalactopyranoside for 5 to 6 h at 22°C. .. Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

Immunohistofluorescence:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: Immunohistofluorescence analysis was carried out on fixed cells and cryostat sections of inner ears, as described ( ). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Infection:

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: Metabolic labeling using PPARα activator Wy-14,643 (100 μ m ) or fenofibrate (100 μ m ) was performed in HeLa cells, and primary hepatocytes were infected with Ad/Med1. .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

other:

Article Title: The G2/M Regulator Histone Demethylase PHF8 Is Targeted for Degradation by the Anaphase-Promoting Complex Containing CDC20
Article Snippet: Antibodies used in this work include anti-PHF8 (catalog numbers ab36068 [Abcam] and A201-772A [Bethyl Laboratories]), anti-RNA polymerase II (Pol II) (CTD4H8) (catalog number 05-623; Millipore), anti-H3 (catalog number 39163; Active Motif), anti-H3K4me3 (MC315) (catalog number 04-745; Millipore), anti-H3K4me2 (CMA303) (catalog number 05-1338; Millipore), anti-H3K9me2 (catalog number ab1220; Abcam), anti-H3K9me1 (catalog number ab8896; Abcam), anti-H3K36me3 (catalog number ab9050; Abcam), anti-H4 (catalog number 39269; Active Motif), anti-H4K20me1 (catalog number ab9051; Abcam), anti-CDC27 (catalog number sc-13154; Santa Cruz), anti-CDC20 (catalog number sc-13162; Santa Cruz), anti-CDH1 (catalog number sc-56381; Santa Cruz), anti-cyclin B1 (catalog number sc-53236; Santa Cruz), anti-cyclin E (catalog number sc-198; Santa Cruz), antiactin (catalog number A2228, Sigma), anti-Flag (M2) (catalog number F1804; Sigma), anti-HA (catalog number MMS-101P; Covance), anti-MYC (catalog number sc-40; Santa Cruz), and anti-HIS (catalog number sc-8036; Santa Cruz).

Article Title: Cholesterol Depletion Reduces Entry of Campylobacter jejuni Cytolethal Distending Toxin and Attenuates Intoxication of Host Cells ▿
Article Snippet: Anti-His (His probe) and anti-proliferating cell nuclear antigen (anti-PCNA) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Article Title: The adenovirus E4-ORF3 protein functions as a SUMO E3 ligase for TIF-1γ sumoylation and poly-SUMO chain elongation
Article Snippet: The anti-HIS (H-15), anti-GST (Z-5), and anti-RanGAP1 (C-5) antibodies were purchased from Santa Cruz Biotechnology.

Article Title: Nuclear translocation of annexin 1 following oxygen-glucose deprivation–reperfusion induces apoptosis by regulating Bid expression via p53 binding
Article Snippet: The following antibodies were used: anti-annexin 1 (Santa Cruz, Dallas, TX, USA; sc-11387, 1 : 1000), anti-histone H3 (Cell Signaling Technology, Beverly, MA, USA; no. 4499, 1 : 1000), anti-p53 (Santa Cruz; sc-126, 1 : 1000), anti-cleaved caspase-3 (Cell Signaling Technology; no. 9664, 1 : 1000), anti-cleaved PARP (Cell Signaling Technology; no. 5625, 1 : 1000), anti-GFP (Santa Cruz; sc-9996, 1 : 1000), anti-His (Santa Cruz; sc-803, 1 : 1000), anti-α -tubulin (Santa Cruz; sc-53646, 1 : 1000), and anti-β -actin (Santa Cruz; sc-47778, 1 : 1000).

Imaging:

Article Title: The Proline-rich N-terminal Domain of G18 Exhibits a Novel G Protein Regulatory Function *
Article Snippet: Membranes were incubated with blocking buffer (Tris-buffered saline Tween 20 (TBST) with 5% skim milk) for 1 h and then probed with anti-His or anti-GST antibody (1:1000) (Santa Cruz biotechnology) diluted in blocking buffer overnight on a rotating platform at 4 °C. .. After another three washes with TBST, the blot was visualized by LumiGLO Reserve chemiluminescence substrate (KPL, Inc.) using a FluorChem 8000 imaging system.

Polymerase Chain Reaction:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. The His-tagged expression vectors of full-length and domain-truncated mutants of hCaMKIINα (as illustrated in ), including pKIINα, pKIINα1–41 , pKIINα1–53 , pKIINα1–68 , and the FLAG-tagged expression vectors of CaMKIIα (pFLAG-CaMKII), and CaMKIIα with H282 mutated to R (H282R) were constructed by PCR cloning and PCR mutation.

Article Title: Establishment of active chromatin structure at enhancer elements by mixed-lineage leukemia 1 to initiate estrogen-dependent gene expression
Article Snippet: Biotinylated PCR products were immobilized on streptavidin-agarose beads and incubated with MLL1 proteins. .. Immunoblotting was performed as described previously , using the following antibodies: anti-His (Santa Cruz Biotechnology); anti-FLAG (Sigma Aldrich); anti-ERα, anti-tubulin (Santa Cruz Biotechnology); anti-MLL1 (Bethyl Laboratories); and anti-FOXA1 (Abcam).

Affinity Purification:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: For immunoelectron microscopy, cochleas from P3 mice (CD1 strain) were labelled with affinity-purified cad-N anti-cadherin 23 antibodies or, as a control, non-immune rabbit IgG ( ). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Binding Assay:

Article Title: Modulation of Rho Guanine Exchange Factor Lfc Activity by Protein Kinase A-Mediated Phosphorylation ▿
Article Snippet: Western blotting and immunofluorescence were performed using the following primary antibodies: anti-FLAG (M2; F3165; Sigma), anti-His (H15; sc-803; Santa Cruz Biotechnology), anti-glutathione S -transferase (anti-GST; B-14; sc-138; Santa Cruz Biotechnology), anti-Gα12 (sc-409; Santa Cruz Biotechnology), anti-Myc (9E10; sc-40; Santa Cruz Biotechnology), anti-GM130 (610822; Becton Dickinson), anti-Myc (610990; Cell Signaling), and antihemagglutinin (anti-HA; H6908; Sigma). .. Anti-14-3-3β (sc-1657; Santa Cruz), anti-AKAP121 (611573), and anti-phospho-Ser-14-3-3 binding motif (4E2; 9606; Cell Signaling Technology) also were used.

Article Title: Establishment of active chromatin structure at enhancer elements by mixed-lineage leukemia 1 to initiate estrogen-dependent gene expression
Article Snippet: DNA pull down assay and immunoblotting FLAG-tagged MLL proteins were expressed in 293T cells, and total cell lysates were used for the binding assay. .. Immunoblotting was performed as described previously , using the following antibodies: anti-His (Santa Cruz Biotechnology); anti-FLAG (Sigma Aldrich); anti-ERα, anti-tubulin (Santa Cruz Biotechnology); anti-MLL1 (Bethyl Laboratories); and anti-FOXA1 (Abcam).

Immunofluorescence:

Article Title: Modulation of Rho Guanine Exchange Factor Lfc Activity by Protein Kinase A-Mediated Phosphorylation ▿
Article Snippet: .. Western blotting and immunofluorescence were performed using the following primary antibodies: anti-FLAG (M2; F3165; Sigma), anti-His (H15; sc-803; Santa Cruz Biotechnology), anti-glutathione S -transferase (anti-GST; B-14; sc-138; Santa Cruz Biotechnology), anti-Gα12 (sc-409; Santa Cruz Biotechnology), anti-Myc (9E10; sc-40; Santa Cruz Biotechnology), anti-GM130 (610822; Becton Dickinson), anti-Myc (610990; Cell Signaling), and antihemagglutinin (anti-HA; H6908; Sigma). .. Horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit secondary antibodies were from Amersham Pharmacia Biotech.

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: Paragraph title: Immunofluorescence and electron microscopy analysis ... The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Molecular Cloning:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. Molecular Cloning and Vector Construction —The full-length hCaMKIINα cDNA was assembled in silico by searching the NCBI data base and amplified by reverse transcription-PCR from bone marrow stromal cells.

In Vivo:

Article Title: Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C]Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C] [W]
Article Snippet: Paragraph title: In Vivo Assays ... Anti-His (catalog no. SC-803; Santa Cruz Biotechnology), anti-Myc (catalog no. SC-40; Santa Cruz Biotechnology), and anti- (catalog no. A190-108A; Bethyl Laboratories) antibodies were used for immunoprecipitation and immunoblotting analyses according to the manufacturers’ protocols.

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: Paragraph title: AMPK in Vitro and in Vivo Phosphorylation Analyses ... After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Pull Down Assay:

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: Paragraph title: In Vitro Pull-Down Assay ... Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

Article Title: Establishment of active chromatin structure at enhancer elements by mixed-lineage leukemia 1 to initiate estrogen-dependent gene expression
Article Snippet: Paragraph title: DNA pull down assay and immunoblotting ... Immunoblotting was performed as described previously , using the following antibodies: anti-His (Santa Cruz Biotechnology); anti-FLAG (Sigma Aldrich); anti-ERα, anti-tubulin (Santa Cruz Biotechnology); anti-MLL1 (Bethyl Laboratories); and anti-FOXA1 (Abcam).

Mutagenesis:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. The His-tagged expression vectors of full-length and domain-truncated mutants of hCaMKIINα (as illustrated in ), including pKIINα, pKIINα1–41 , pKIINα1–53 , pKIINα1–68 , and the FLAG-tagged expression vectors of CaMKIIα (pFLAG-CaMKII), and CaMKIIα with H282 mutated to R (H282R) were constructed by PCR cloning and PCR mutation.

Article Title: Establishment of active chromatin structure at enhancer elements by mixed-lineage leukemia 1 to initiate estrogen-dependent gene expression
Article Snippet: Point mutations in MLL1 were introduced by QuikChange site-directed mutagenesis kit (Stratagene). .. Immunoblotting was performed as described previously , using the following antibodies: anti-His (Santa Cruz Biotechnology); anti-FLAG (Sigma Aldrich); anti-ERα, anti-tubulin (Santa Cruz Biotechnology); anti-MLL1 (Bethyl Laboratories); and anti-FOXA1 (Abcam).

Immuno-Electron Microscopy:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: For immunoelectron microscopy, cochleas from P3 mice (CD1 strain) were labelled with affinity-purified cad-N anti-cadherin 23 antibodies or, as a control, non-immune rabbit IgG ( ). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Immunohistochemistry:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. An hCaMKIINα-specific polyclonal antibody was raised against affinity chromatography-purified GST-KIINα1–41 fusion protein, and the antibody specificity was confirmed by Western blot and immunohistochemical staining for no cross-reactivity with hCaMKIINβ.

Labeling:

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: After metabolic labeling, cells were washed once in ice-cold phosphate-free buffer before lysis in ice-cold radioimmune precipitation assay buffer containing 1% (w/w) Nonidet P-40, 1% (w/v) sodium deoxycholate, 0.1% (w/v) SDS, 0.15 m NaCl, 0.01 m sodium phosphate (pH 7.2), 2 m m EDTA, 50 m m NaF, 1 m m EGTA, 1 m m PMSF, 0.2 m m sodium orthovanadate, and 0.1 mg/ml protease inhibitor mixture. .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Mouse Assay:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: For immunoelectron microscopy, cochleas from P3 mice (CD1 strain) were labelled with affinity-purified cad-N anti-cadherin 23 antibodies or, as a control, non-immune rabbit IgG ( ). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Microscopy:

Article Title: Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair cell bundle
Article Snippet: Cells and tissue sections were analysed with a laser scanning confocal microscope (LSM-510, Zeiss). .. The following mouse monoclonal antibodies were used: anti-Myc (clone 9E10) and anti-His (Santa Cruz); anti-GST (Amersham); anti-vinculin (Sigma); and anti-V5 (Invitrogen).

Staining:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. An hCaMKIINα-specific polyclonal antibody was raised against affinity chromatography-purified GST-KIINα1–41 fusion protein, and the antibody specificity was confirmed by Western blot and immunohistochemical staining for no cross-reactivity with hCaMKIINβ.

Purification:

Article Title: Establishment of active chromatin structure at enhancer elements by mixed-lineage leukemia 1 to initiate estrogen-dependent gene expression
Article Snippet: The 6 × His-tagged proteins were expressed in Escherichia coli strain BL21(DE3), purified by incubation with Ni-NTA agarose beads and eluted with phosphate-buffered saline (PBS) containing 250 mM imidazole. .. Immunoblotting was performed as described previously , using the following antibodies: anti-His (Santa Cruz Biotechnology); anti-FLAG (Sigma Aldrich); anti-ERα, anti-tubulin (Santa Cruz Biotechnology); anti-MLL1 (Bethyl Laboratories); and anti-FOXA1 (Abcam).

Chromatin Immunoprecipitation:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: Paragraph title: Chromatin immunoprecipitation (ChIP) assay ... They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.).

SDS Page:

Article Title: Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C]Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C] [W]
Article Snippet: The clear supernatant was subjected to 12% SDS-PAGE followed by immunoblot analysis. .. Anti-His (catalog no. SC-803; Santa Cruz Biotechnology), anti-Myc (catalog no. SC-40; Santa Cruz Biotechnology), and anti- (catalog no. A190-108A; Bethyl Laboratories) antibodies were used for immunoprecipitation and immunoblotting analyses according to the manufacturers’ protocols.

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: .. Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody. .. Two hundred milliliters of E. coli cell culture expressing His-CBP1 (0.5 mM isopropyl β- d -1-thiogalactopyranoside, 16°C, overnight) was centrifuged at 8000 rpm at 4°C for 2 min, and the pellet was resuspended in 8 mL cold lysis buffer (50 mM NaH2 PO4 , 300 mM NaCl, 20 mM imidazole, and 1× proteinase inhibitor cocktail [Roche], pH 8.0) followed by sonication on ice (3-s bursts with 3-s cooling periods, 15 min in total).

Article Title: Functional Characterization of EscK (Orf4), a Sorting Platform Component of the Enteropathogenic Escherichia coli Injectisome
Article Snippet: Protein samples were subjected to 12 or 15% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) or nitrocellulose membranes. .. The commercial antibodies anti-DnaK (MBL International), horseradish peroxidase (HRP)-conjugated anti-HA (Roche), anti-MBP (New England BioLabs), anti-His (Santa Cruz Biotechnology), and anti-FLAG (Sigma) were used according to the manufacturers' instructions.

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare). .. After washing with Tris-buffered saline, the protein was eluted in Laemmli sample buffer, subjected to SDS-PAGE, vacuum-dried, and autoradiographed.

Plasmid Preparation:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: Briefly, 5×106 HCT116/MMTV cells grown in 15 cm-dishes were transfected with 1 μg/well of plasmid expressing the His-tagged full-length, Δ181–225, 181–225 fragment of Set/TAF-Iβ or the His-tagged Set-Can, together with 10 μg/well of pRShGRα. .. They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.).

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. Molecular Cloning and Vector Construction —The full-length hCaMKIINα cDNA was assembled in silico by searching the NCBI data base and amplified by reverse transcription-PCR from bone marrow stromal cells.

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: For AMPK-dependent in vivo phosphorylation of Med1, HEK293 cells, HeLa cells, and mouse primary hepatocytes were infected with Ad-Med1 (3.5 × 106 virus particle) or transiently transfected with 24 μg of pShuttle-His-Med1 plasmid using Lipofectamine (Invitrogen). .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Functional Assay:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.). .. The promoter region -219 to -47 of the MMTV long terminal repeat (fragment size 173 bps), which contains two functional GREs, was amplified from the prepared DNA samples using a primer pair: 5′-AACCTTGCGGTTCCCAG-3′ and 5′-GCATTTACATAAGATTTGG-3′, while tandem endogenous GREs of the rat TAT promoter, which are located ~2,500 bps upstream of its transcription initiation site, were amplified by a primer pair: 5′-TCTTCTCAGTGTTCTCTATCAC-3′ and 5′-CAGAAACCGACAGGCGACTACG-3′ (fragment size 220 bps), as described previously [ , ].

Agarose Gel Electrophoresis:

Article Title: Activated Glucocorticoid Receptor Interacts with the INHAT Component Set/TAF-I? and Releases it from a Glucocorticoid-responsive Gene Promoter, Relieving Repression: Implications for the Pathogenesis of Glucocorticoid Resistance in Acute Undifferentiated Leukemia with Set-Can Translocation
Article Snippet: They were then fixed with 1% formaldehyde and homogenized, and cross-linked DNA/protein complexes were co-precipitated with anti-His, anti-Set/TAF-Iβ, anti-GRα, anti-GRIP1 (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA) or anti-pp32 (IMEGENEX, San Diego, CA) antibodies, or rabbit control IgG (Santa Cruz Biotechnologies, Inc.). .. Amplified products were then run on a 2–3% agarose gel, and visualized DNA bands were photographed.

In Vitro:

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: Paragraph title: In Vitro Pull-Down Assay ... Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: Paragraph title: AMPK in Vitro and in Vivo Phosphorylation Analyses ... After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Immunoprecipitation:

Article Title: Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C]Rice Mitogen-Activated Protein Kinase Interactome Analysis Using the Yeast Two-Hybrid System 1 [C] [W]
Article Snippet: .. Anti-His (catalog no. SC-803; Santa Cruz Biotechnology), anti-Myc (catalog no. SC-40; Santa Cruz Biotechnology), and anti- (catalog no. A190-108A; Bethyl Laboratories) antibodies were used for immunoprecipitation and immunoblotting analyses according to the manufacturers’ protocols. .. The protein expression vectors used were pRSET A (Invitrogen) and pGEX4T-1 (Amersham Biosciences).

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare). .. After washing with Tris-buffered saline, the protein was eluted in Laemmli sample buffer, subjected to SDS-PAGE, vacuum-dried, and autoradiographed.

Construct:

Article Title: A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization *A Novel Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest via p27 Stabilization * S⃞
Article Snippet: Anti-His, p27, phosphorylated p27(T187), and MEK1 were from Santa Cruz Biotechnology, Santa Cruz, CA); anti-ERK1/2, phosphorylated MEK1, and phosphorylated ERK1/2 antibodies were from Cell Signaling Technology; and anti-FLAG, actin, and CaMKIIα antibodies were from Sigma. .. The His-tagged expression vectors of full-length and domain-truncated mutants of hCaMKIINα (as illustrated in ), including pKIINα, pKIINα1–41 , pKIINα1–53 , pKIINα1–68 , and the FLAG-tagged expression vectors of CaMKIIα (pFLAG-CaMKII), and CaMKIIα with H282 mutated to R (H282R) were constructed by PCR cloning and PCR mutation.

Article Title: Arabidopsis CBP1 Is a Novel Regulator of Transcription Initiation in Central Cell-Mediated Pollen Tube Guidance [OPEN]
Article Snippet: The fusion constructs were transformed into Escherichia coli strain BL21. .. Finally, the proteins bound on the beads were boiled with 1× SDS loading buffer in a 95 to 100°C water bath and then subjected to SDS-PAGE and immunoblot with anti-GST (GE) and anti-His (Santa Cruz) antibody.

Protease Inhibitor:

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: After metabolic labeling, cells were washed once in ice-cold phosphate-free buffer before lysis in ice-cold radioimmune precipitation assay buffer containing 1% (w/w) Nonidet P-40, 1% (w/v) sodium deoxycholate, 0.1% (w/v) SDS, 0.15 m NaCl, 0.01 m sodium phosphate (pH 7.2), 2 m m EDTA, 50 m m NaF, 1 m m EGTA, 1 m m PMSF, 0.2 m m sodium orthovanadate, and 0.1 mg/ml protease inhibitor mixture. .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

Lysis:

Article Title: The Med1 Subunit of the Mediator Complex Induces Liver Cell Proliferation and Is Phosphorylated by AMP Kinase *
Article Snippet: After metabolic labeling, cells were washed once in ice-cold phosphate-free buffer before lysis in ice-cold radioimmune precipitation assay buffer containing 1% (w/w) Nonidet P-40, 1% (w/v) sodium deoxycholate, 0.1% (w/v) SDS, 0.15 m NaCl, 0.01 m sodium phosphate (pH 7.2), 2 m m EDTA, 50 m m NaF, 1 m m EGTA, 1 m m PMSF, 0.2 m m sodium orthovanadate, and 0.1 mg/ml protease inhibitor mixture. .. After lysates were cleared by high speed centrifugation (26,000 × g for 30 min), His-Med1 was immunoprecipitated using either anti-Med1 (4 μg; Santa Cruz Biotechnology, sc8998) or anti-His (4 μg; Santa Cruz Biotechnology, sc-803) conjugated with protein G beads (GE Healthcare).

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  • 86
    Santa Cruz Biotechnology fitc labeled anti his tag monoclonal antibody mab
    Immuno-fluorescence staining detection for the receptor-binding of the fusion protein. A375.S2 cells incubated with 100 μg/ml fusion protein ( 1 ), with 10 μg/ml fusion protein ( 2 ), with 1 μg/ml fusion protein ( 3 ), with 0.1 μg/ml fusion protein ( 4 ), with 0 μg/ml fusion protein ( 5 ), and with 100 μg/ml HSA ( 6 ). <t>FITC-labeled</t> anti-His tag <t>mAb</t> is used in this assay. The fusion protein, IL-1ra and HSA have a His tag at the C-terminal.
    Fitc Labeled Anti His Tag Monoclonal Antibody Mab, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc labeled anti his tag monoclonal antibody mab/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fitc labeled anti his tag monoclonal antibody mab - by Bioz Stars, 2020-04
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    84
    Santa Cruz Biotechnology rabbit polyclonal anti his tag
    Fig. 2. Interaction of LZIP with HCV core protein. ( A ). ( B ) Co-immunoprecipitation of LZIP and core protein from transfected cells. HeLa and HepG2 cells were transfected individually with the plasmids indicated (5 μg). Immunoprecipitation was performed with cleared cell lysates using a mouse monoclonal anti-Gal4BD antibody [α-Gal4 (m)]. Blots were probed with rabbit <t>polyclonal</t> anti-core [α-C (r); lanes 1–4] or anti-His tag [α-His (r); lanes 5–8]. Bands representing core (C) or His-tagged LZIP (LZIP) co-precipitated by α-Gal4 are indicated. pM is an expression vector for Gal4BD. ( C ) LZIP binds to immobilized GST–C but not to GST. His-tagged LZIP (5 μg) was loaded onto either GST- or GST–C–bound glutathione–Sepharose (Pharmacia). The resins were washed extensively. Proteins bound to the resins were solubilized in SDS gel loading buffer, resolved by 10% SDS–PAGE, transferred to membrane and probed with anti-GST (lanes 1 and 2; α-GST), anti-His tag (lanes 3 and 4; α-His) or anti-LZIP (lanes 5 and 6; α-ZN). Lanes 1 and 2 show the immobilized GST and GST–C proteins in the resins. His-tagged LZIP was retained in the GST–C resin (lanes 4 and 6) and not in the GST resin (lanes 3 and 5). We verified that ∼1 μg of the His-tagged LZIP loaded onto the GST–C resin (∼20% of the input) bound to GST–C.
    Rabbit Polyclonal Anti His Tag, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 84/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti his tag/product/Santa Cruz Biotechnology
    Average 84 stars, based on 4 article reviews
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    87
    Santa Cruz Biotechnology hrp conjugated anti his tag antibody
    ELISA of binding of <t>IgG</t> and Fc fusion proteins to immobilized CEA. Bound proteins were detected with an anti-Fc <t>HRP-conjugated</t> antibody.
    Hrp Conjugated Anti His Tag Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated anti his tag antibody/product/Santa Cruz Biotechnology
    Average 87 stars, based on 1 article reviews
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    Image Search Results


    Immuno-fluorescence staining detection for the receptor-binding of the fusion protein. A375.S2 cells incubated with 100 μg/ml fusion protein ( 1 ), with 10 μg/ml fusion protein ( 2 ), with 1 μg/ml fusion protein ( 3 ), with 0.1 μg/ml fusion protein ( 4 ), with 0 μg/ml fusion protein ( 5 ), and with 100 μg/ml HSA ( 6 ). FITC-labeled anti-His tag mAb is used in this assay. The fusion protein, IL-1ra and HSA have a His tag at the C-terminal.

    Journal: BMC Biotechnology

    Article Title: Selective delivery of interleukine-1 receptor antagonist to inflamed joint by albumin fusion

    doi: 10.1186/1472-6750-12-68

    Figure Lengend Snippet: Immuno-fluorescence staining detection for the receptor-binding of the fusion protein. A375.S2 cells incubated with 100 μg/ml fusion protein ( 1 ), with 10 μg/ml fusion protein ( 2 ), with 1 μg/ml fusion protein ( 3 ), with 0.1 μg/ml fusion protein ( 4 ), with 0 μg/ml fusion protein ( 5 ), and with 100 μg/ml HSA ( 6 ). FITC-labeled anti-His tag mAb is used in this assay. The fusion protein, IL-1ra and HSA have a His tag at the C-terminal.

    Article Snippet: FITC-labeled anti-His tag monoclonal antibody (mAb) and HRP-conjugated anti-His tag mAb were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).Restriction endonuclease, T4 DNA ligase, DNase, and RNase were from TaKaRa Biotechnology (Dalian, China).

    Techniques: Fluorescence, Staining, Binding Assay, Incubation, Labeling

    Fig. 2. Interaction of LZIP with HCV core protein. ( A ). ( B ) Co-immunoprecipitation of LZIP and core protein from transfected cells. HeLa and HepG2 cells were transfected individually with the plasmids indicated (5 μg). Immunoprecipitation was performed with cleared cell lysates using a mouse monoclonal anti-Gal4BD antibody [α-Gal4 (m)]. Blots were probed with rabbit polyclonal anti-core [α-C (r); lanes 1–4] or anti-His tag [α-His (r); lanes 5–8]. Bands representing core (C) or His-tagged LZIP (LZIP) co-precipitated by α-Gal4 are indicated. pM is an expression vector for Gal4BD. ( C ) LZIP binds to immobilized GST–C but not to GST. His-tagged LZIP (5 μg) was loaded onto either GST- or GST–C–bound glutathione–Sepharose (Pharmacia). The resins were washed extensively. Proteins bound to the resins were solubilized in SDS gel loading buffer, resolved by 10% SDS–PAGE, transferred to membrane and probed with anti-GST (lanes 1 and 2; α-GST), anti-His tag (lanes 3 and 4; α-His) or anti-LZIP (lanes 5 and 6; α-ZN). Lanes 1 and 2 show the immobilized GST and GST–C proteins in the resins. His-tagged LZIP was retained in the GST–C resin (lanes 4 and 6) and not in the GST resin (lanes 3 and 5). We verified that ∼1 μg of the His-tagged LZIP loaded onto the GST–C resin (∼20% of the input) bound to GST–C.

    Journal: The EMBO Journal

    Article Title: Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation

    doi: 10.1093/emboj/19.4.729

    Figure Lengend Snippet: Fig. 2. Interaction of LZIP with HCV core protein. ( A ). ( B ) Co-immunoprecipitation of LZIP and core protein from transfected cells. HeLa and HepG2 cells were transfected individually with the plasmids indicated (5 μg). Immunoprecipitation was performed with cleared cell lysates using a mouse monoclonal anti-Gal4BD antibody [α-Gal4 (m)]. Blots were probed with rabbit polyclonal anti-core [α-C (r); lanes 1–4] or anti-His tag [α-His (r); lanes 5–8]. Bands representing core (C) or His-tagged LZIP (LZIP) co-precipitated by α-Gal4 are indicated. pM is an expression vector for Gal4BD. ( C ) LZIP binds to immobilized GST–C but not to GST. His-tagged LZIP (5 μg) was loaded onto either GST- or GST–C–bound glutathione–Sepharose (Pharmacia). The resins were washed extensively. Proteins bound to the resins were solubilized in SDS gel loading buffer, resolved by 10% SDS–PAGE, transferred to membrane and probed with anti-GST (lanes 1 and 2; α-GST), anti-His tag (lanes 3 and 4; α-His) or anti-LZIP (lanes 5 and 6; α-ZN). Lanes 1 and 2 show the immobilized GST and GST–C proteins in the resins. His-tagged LZIP was retained in the GST–C resin (lanes 4 and 6) and not in the GST resin (lanes 3 and 5). We verified that ∼1 μg of the His-tagged LZIP loaded onto the GST–C resin (∼20% of the input) bound to GST–C.

    Article Snippet: Rabbit polyclonal anti-His tag (H-15) and mouse monoclonal anti-Gal4BD (clone RK5C1) were from Santa Cruz.

    Techniques: Immunoprecipitation, Transfection, Expressing, Plasmid Preparation, SDS-Gel, SDS Page

    (A) Analysis of actin distribution and length of flagellar axoneme in LdCof, S4D, and S4A protein-overexpressing cells. In wild-type and overexpressing cells, actin and tubulin were stained with polyclonal rabbit anti- Leishmania actin antibodies and polyclonal

    Journal: Eukaryotic Cell

    Article Title: Overexpression of S4D Mutant of Leishmania donovani ADF/Cofilin Impairs Flagellum Assembly by Affecting Actin Dynamics

    doi: 10.1128/EC.00013-12

    Figure Lengend Snippet: (A) Analysis of actin distribution and length of flagellar axoneme in LdCof, S4D, and S4A protein-overexpressing cells. In wild-type and overexpressing cells, actin and tubulin were stained with polyclonal rabbit anti- Leishmania actin antibodies and polyclonal

    Article Snippet: Anti-His rabbit polyclonal antibodies were obtained from Santa Cruz Biotechnology.

    Techniques: Staining

    (A) SDS-PAGE analysis showing overexpression of LdCof, S4D, and S4A proteins in Leishmania donovani cells. Panel a, Coomassie blue-stained 12% SDS-PAGE of total cell lysates showing equal loading of samples. Panel b, Western blot of panel a using polyclonal

    Journal: Eukaryotic Cell

    Article Title: Overexpression of S4D Mutant of Leishmania donovani ADF/Cofilin Impairs Flagellum Assembly by Affecting Actin Dynamics

    doi: 10.1128/EC.00013-12

    Figure Lengend Snippet: (A) SDS-PAGE analysis showing overexpression of LdCof, S4D, and S4A proteins in Leishmania donovani cells. Panel a, Coomassie blue-stained 12% SDS-PAGE of total cell lysates showing equal loading of samples. Panel b, Western blot of panel a using polyclonal

    Article Snippet: Anti-His rabbit polyclonal antibodies were obtained from Santa Cruz Biotechnology.

    Techniques: SDS Page, Over Expression, Staining, Western Blot

    ELISA of binding of IgG and Fc fusion proteins to immobilized CEA. Bound proteins were detected with an anti-Fc HRP-conjugated antibody.

    Journal: mAbs

    Article Title: Pharmacokinetic properties of IgG and various Fc fusion proteins in mice

    doi: 10.1080/19420862.2015.1113360

    Figure Lengend Snippet: ELISA of binding of IgG and Fc fusion proteins to immobilized CEA. Bound proteins were detected with an anti-Fc HRP-conjugated antibody.

    Article Snippet: Detection was performed with HRP-conjugated anti-His-tag antibody (scDb-CH2) or anti-human IgG (Fc-specific) antibody using 100 µl 3,3′,5,5′tetramethylbenzidine (TMB) substrate (0.1 mg/ml TMB, 100 mM sodium acetate buffer pH 6, 0.006% H2 O2 ).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay