Structured Review

GE Healthcare pbs t
T β R-II protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in <t>PBS-T,</t> and incubated with T β R-II antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 70-kDa band corresponding to the size expected for T β R-II protein was observed in all cell lines in total lysate and the cytosolic fraction. The membrane fractions demonstrate the clear presence of T β R-II protein in the Mv1Lu positive control, the MCF-10F nontumorigenic, and the tumorigenic MDA-MB231 cell lines. The other cell lines have little or no T β R-II protein present in the membrane fraction.
Pbs T, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pbs t/product/GE Healthcare
Average 92 stars, based on 48 article reviews
Price from $9.99 to $1999.99
pbs t - by Bioz Stars, 2020-07
92/100 stars

Images

1) Product Images from "Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells"

Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells

Journal: Gene Expression

doi:

T β R-II protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-II antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 70-kDa band corresponding to the size expected for T β R-II protein was observed in all cell lines in total lysate and the cytosolic fraction. The membrane fractions demonstrate the clear presence of T β R-II protein in the Mv1Lu positive control, the MCF-10F nontumorigenic, and the tumorigenic MDA-MB231 cell lines. The other cell lines have little or no T β R-II protein present in the membrane fraction.
Figure Legend Snippet: T β R-II protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-II antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 70-kDa band corresponding to the size expected for T β R-II protein was observed in all cell lines in total lysate and the cytosolic fraction. The membrane fractions demonstrate the clear presence of T β R-II protein in the Mv1Lu positive control, the MCF-10F nontumorigenic, and the tumorigenic MDA-MB231 cell lines. The other cell lines have little or no T β R-II protein present in the membrane fraction.

Techniques Used: Expressing, SDS Page, Electrophoresis, Incubation, Autoradiography, Positive Control, Multiple Displacement Amplification

T β R-I protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-I antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 55-kDa band corresponding to the presence of T β R-I protein in total lysate, cytosolic, and membrane-bound protein fractions was observed. Membrane-bound fractions (and to a lesser extent the total cellular lysate) contain two (or three in the case of MCF-7) bands of close molecular weight, the significance of which remains to be determined.
Figure Legend Snippet: T β R-I protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-I antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 55-kDa band corresponding to the presence of T β R-I protein in total lysate, cytosolic, and membrane-bound protein fractions was observed. Membrane-bound fractions (and to a lesser extent the total cellular lysate) contain two (or three in the case of MCF-7) bands of close molecular weight, the significance of which remains to be determined.

Techniques Used: Expressing, SDS Page, Electrophoresis, Incubation, Autoradiography, Molecular Weight

2) Product Images from "A compact phage display human scFv library for selection of antibodies to a wide variety of antigens"

Article Title: A compact phage display human scFv library for selection of antibodies to a wide variety of antigens

Journal: BMC Biotechnology

doi: 10.1186/1472-6750-9-6

Inhibition ELISA of anti-Aflatoxin B1 antibody . Various 1:2 dilutions of soluble Aflatoxin B1 from 5.0–0.039 μg/ml in 2% MPBS were incubated with three selected Phage (C3, C5, D2) at 37°C for 30 min before adding into wells of Immuno 96 MicroWell™ Plates, coated with 4 μg/ml BSA-conjugated AlatoxinB1. After 1 hr of incubation, the plates were washed three times with PBS-T followed by 3 times with PBS. Bound phage were detected with anti-M13 phage-horseradish peroxidase (HRP) conjugate, using ABTS (2,2-azino-di-3-ethyl-benzthiazoine-6-sulfonate) as a substrate. The average absorbance at a wavelength of 405 nm and S.D. are shown.
Figure Legend Snippet: Inhibition ELISA of anti-Aflatoxin B1 antibody . Various 1:2 dilutions of soluble Aflatoxin B1 from 5.0–0.039 μg/ml in 2% MPBS were incubated with three selected Phage (C3, C5, D2) at 37°C for 30 min before adding into wells of Immuno 96 MicroWell™ Plates, coated with 4 μg/ml BSA-conjugated AlatoxinB1. After 1 hr of incubation, the plates were washed three times with PBS-T followed by 3 times with PBS. Bound phage were detected with anti-M13 phage-horseradish peroxidase (HRP) conjugate, using ABTS (2,2-azino-di-3-ethyl-benzthiazoine-6-sulfonate) as a substrate. The average absorbance at a wavelength of 405 nm and S.D. are shown.

Techniques Used: Inhibition, Enzyme-linked Immunosorbent Assay, Incubation

3) Product Images from "Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells"

Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells

Journal: Gene Expression

doi:

T β R-II protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-II antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 70-kDa band corresponding to the size expected for T β R-II protein was observed in all cell lines in total lysate and the cytosolic fraction. The membrane fractions demonstrate the clear presence of T β R-II protein in the Mv1Lu positive control, the MCF-10F nontumorigenic, and the tumorigenic MDA-MB231 cell lines. The other cell lines have little or no T β R-II protein present in the membrane fraction.
Figure Legend Snippet: T β R-II protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-II antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 70-kDa band corresponding to the size expected for T β R-II protein was observed in all cell lines in total lysate and the cytosolic fraction. The membrane fractions demonstrate the clear presence of T β R-II protein in the Mv1Lu positive control, the MCF-10F nontumorigenic, and the tumorigenic MDA-MB231 cell lines. The other cell lines have little or no T β R-II protein present in the membrane fraction.

Techniques Used: Expressing, SDS Page, Electrophoresis, Incubation, Autoradiography, Positive Control, Multiple Displacement Amplification

T β R-I protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-I antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 55-kDa band corresponding to the presence of T β R-I protein in total lysate, cytosolic, and membrane-bound protein fractions was observed. Membrane-bound fractions (and to a lesser extent the total cellular lysate) contain two (or three in the case of MCF-7) bands of close molecular weight, the significance of which remains to be determined.
Figure Legend Snippet: T β R-I protein expression in total lysate, cytosol, and membrane fractions from human breast cell lines. Aliquots (10 μg) of total cell lysate and fractionated protein were analyzed by SDS-PAGE electrophoresis, transferred to PVDF membranes, blocked with 5% nonfat dry milk in PBS-T, and incubated with T β R-I antibody. Bound antibodies were detected by incubation with peroxidase-conjugated antirabbit immunoglobulin and enhanced chemiluminscent autoradiography. A 55-kDa band corresponding to the presence of T β R-I protein in total lysate, cytosolic, and membrane-bound protein fractions was observed. Membrane-bound fractions (and to a lesser extent the total cellular lysate) contain two (or three in the case of MCF-7) bands of close molecular weight, the significance of which remains to be determined.

Techniques Used: Expressing, SDS Page, Electrophoresis, Incubation, Autoradiography, Molecular Weight

4) Product Images from "Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice"

Article Title: Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice

Journal: Comparative Hepatology

doi: 10.1186/1476-5926-11-1

Immunohistochemical staining of liver sections for Abcc3 detection. Frozen livers were cut to 5 μm cryosections and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS). Sections were blocked in goat serum followed by incubation with anti -Abcc3. Sections were washed with PBS and incubated with goat anti -rat IgG conjugated to Alexfluor 488 (green staining) and rhodamine-conjugated phalloidin (red). Sections were then rinsed with PBS/T, PBS, and water, air dried, and then mounted with Prolong Gold containing DAPI (blue staining). All images are displayed as 200X magnification. It was observed that green staining displaying Abcc3 expression was higher in db/db male and female mice as compared to controls.
Figure Legend Snippet: Immunohistochemical staining of liver sections for Abcc3 detection. Frozen livers were cut to 5 μm cryosections and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS). Sections were blocked in goat serum followed by incubation with anti -Abcc3. Sections were washed with PBS and incubated with goat anti -rat IgG conjugated to Alexfluor 488 (green staining) and rhodamine-conjugated phalloidin (red). Sections were then rinsed with PBS/T, PBS, and water, air dried, and then mounted with Prolong Gold containing DAPI (blue staining). All images are displayed as 200X magnification. It was observed that green staining displaying Abcc3 expression was higher in db/db male and female mice as compared to controls.

Techniques Used: Immunohistochemistry, Staining, Incubation, Expressing, Mouse Assay

Related Articles

Fluorescence:

Article Title: Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice
Article Snippet: .. After incubation with secondary antibody, membranes were washed three times in PBS/T, incubated with ECL + fluorescence Reagent (GE Healthcare, Buckinghamshire, UK), and developed using autoradiography. .. Protein bands on autoradiographs were quantified using Quantity One® software v4.6.3 (Biorad, Hercules, CA).

Autoradiography:

Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells
Article Snippet: .. Bound antibodies were detected by incubation with peroxidase-conjugated anti-rabbit immunogloblins (Amersham, Arlington Heights, IL) at 1:5000 dilution in PBS-T for 1 h at room temperature, washing 3 in PBS-T, and enhanced chemiluminscent autoradiography (ECL: Amersham). ..

Article Title: Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice
Article Snippet: .. After incubation with secondary antibody, membranes were washed three times in PBS/T, incubated with ECL + fluorescence Reagent (GE Healthcare, Buckinghamshire, UK), and developed using autoradiography. .. Protein bands on autoradiographs were quantified using Quantity One® software v4.6.3 (Biorad, Hercules, CA).

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Briefly, membranes loaded with oligopeptides (see above) were incubated with 5 to 10 ml of a host cell suspension, i.e., EBL tissue culture cells metabolically labeled with 35 S-methionine (0.74 MBq per 5 ml of culture; Amersham), at 37°C with intensive shaking for 2 h. After three washes in PBS-T, membranes were air dried, and reactive spots were visualized within 1 to 4 days by autoradiography with Hyperfilm-ßmax (Amersham). ..

Labeling:

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Briefly, membranes loaded with oligopeptides (see above) were incubated with 5 to 10 ml of a host cell suspension, i.e., EBL tissue culture cells metabolically labeled with 35 S-methionine (0.74 MBq per 5 ml of culture; Amersham), at 37°C with intensive shaking for 2 h. After three washes in PBS-T, membranes were air dried, and reactive spots were visualized within 1 to 4 days by autoradiography with Hyperfilm-ßmax (Amersham). ..

Incubation:

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Before each trial of epitope analysis, nonspecific active sites were blocked by incubation of the membranes in PBS–0.3% Tween 20 (PBS-T) containing 2% (wt/vol) skim milk powder at room temperature for 30 min. Membranes were incubated with 10 ml of animal serum (1:20) in PBS-T at room temperature for 1 h, and reactive spots were visualized by chemiluminescence (ECL Western Blotting Detection System; Amersham) after 30 min of incubation with peroxidase-labeled anti-bovine immunoglobulin G diluted 1:10,000. .. Membranes were used again after 30 min of incubation at 50°C in stripping buffer (100 mM 2-mercaptoethanol sulfonic acid [sodium salt], 2% [wt/vol] sodium dodecyl sulfate, 62.5 mM Tris [pH 6.7]).

Article Title: Use of Recombinant gp43 Isoforms Expressed in Pichia pastoris for Diagnosis of Paracoccidioidomycosis
Article Snippet: .. Sera were diluted in PBS-M and incubated with shaking for 1 h at room temperature with the membranes, which were then washed three times in PBS-T and incubated with secondary antibodies (peroxidase-labeled goat anti-human immunoglobulin G [IgG] or anti-rabbit immunoglobulin, used at 1:2,000; Amersham Biosciences) for 1 h at room temperature, with shaking. .. The membranes were washed three times, and the reactions were developed with diaminobenzidine (Sigma).

Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells
Article Snippet: .. Bound antibodies were detected by incubation with peroxidase-conjugated anti-rabbit immunogloblins (Amersham, Arlington Heights, IL) at 1:5000 dilution in PBS-T for 1 h at room temperature, washing 3 in PBS-T, and enhanced chemiluminscent autoradiography (ECL: Amersham). ..

Article Title: Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice
Article Snippet: .. After incubation with secondary antibody, membranes were washed three times in PBS/T, incubated with ECL + fluorescence Reagent (GE Healthcare, Buckinghamshire, UK), and developed using autoradiography. .. Protein bands on autoradiographs were quantified using Quantity One® software v4.6.3 (Biorad, Hercules, CA).

Article Title: A compact phage display human scFv library for selection of antibodies to a wide variety of antigens
Article Snippet: .. The plates were washed three times with PBS-T, followed by three times with PBS, and incubated with a 1:5,000 dilution of a mouse anti-M13 phage-horseradish peroxidase (HRP) conjugate (Amersham-Pharmacia Biotech, Sweden) in 2% MPBS. .. The ABTS (2,2-azino-di-3-ethyl-benzthiazoine-6-sulfonate) peroxidase substrate (Fluka, USA) was added, and the absorbance was read at 405 nm, using a Sunrise absorbance reader (TECAN, Austria).

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Briefly, membranes loaded with oligopeptides (see above) were incubated with 5 to 10 ml of a host cell suspension, i.e., EBL tissue culture cells metabolically labeled with 35 S-methionine (0.74 MBq per 5 ml of culture; Amersham), at 37°C with intensive shaking for 2 h. After three washes in PBS-T, membranes were air dried, and reactive spots were visualized within 1 to 4 days by autoradiography with Hyperfilm-ßmax (Amersham). ..

Metabolic Labelling:

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Briefly, membranes loaded with oligopeptides (see above) were incubated with 5 to 10 ml of a host cell suspension, i.e., EBL tissue culture cells metabolically labeled with 35 S-methionine (0.74 MBq per 5 ml of culture; Amersham), at 37°C with intensive shaking for 2 h. After three washes in PBS-T, membranes were air dried, and reactive spots were visualized within 1 to 4 days by autoradiography with Hyperfilm-ßmax (Amersham). ..

Western Blot:

Article Title: Epitope Mapping of Immunogenic and Adhesive Structures in Repetitive Domains of Mycoplasma bovis Variable Surface Lipoproteins
Article Snippet: .. Before each trial of epitope analysis, nonspecific active sites were blocked by incubation of the membranes in PBS–0.3% Tween 20 (PBS-T) containing 2% (wt/vol) skim milk powder at room temperature for 30 min. Membranes were incubated with 10 ml of animal serum (1:20) in PBS-T at room temperature for 1 h, and reactive spots were visualized by chemiluminescence (ECL Western Blotting Detection System; Amersham) after 30 min of incubation with peroxidase-labeled anti-bovine immunoglobulin G diluted 1:10,000. .. Membranes were used again after 30 min of incubation at 50°C in stripping buffer (100 mM 2-mercaptoethanol sulfonic acid [sodium salt], 2% [wt/vol] sodium dodecyl sulfate, 62.5 mM Tris [pH 6.7]).

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    GE Healthcare pbs t
    Immunohistochemical staining of liver sections for Abcc3 detection. Frozen livers were cut to 5 μm cryosections and fixed in 4% paraformaldehyde in phosphate-buffered saline <t>(PBS).</t> Sections were blocked in goat serum followed by incubation with anti -Abcc3. Sections were washed with PBS and incubated with goat anti -rat IgG conjugated to Alexfluor 488 (green staining) and rhodamine-conjugated phalloidin (red). Sections were then rinsed with <t>PBS/T,</t> PBS, and water, air dried, and then mounted with Prolong Gold containing DAPI (blue staining). All images are displayed as 200X magnification. It was observed that green staining displaying Abcc3 expression was higher in db/db male and female mice as compared to controls.
    Pbs T, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs t/product/GE Healthcare
    Average 92 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
    pbs t - by Bioz Stars, 2020-07
    92/100 stars
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    Immunohistochemical staining of liver sections for Abcc3 detection. Frozen livers were cut to 5 μm cryosections and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS). Sections were blocked in goat serum followed by incubation with anti -Abcc3. Sections were washed with PBS and incubated with goat anti -rat IgG conjugated to Alexfluor 488 (green staining) and rhodamine-conjugated phalloidin (red). Sections were then rinsed with PBS/T, PBS, and water, air dried, and then mounted with Prolong Gold containing DAPI (blue staining). All images are displayed as 200X magnification. It was observed that green staining displaying Abcc3 expression was higher in db/db male and female mice as compared to controls.

    Journal: Comparative Hepatology

    Article Title: Severe diabetes and leptin resistance cause differential hepatic and renal transporter expression in mice

    doi: 10.1186/1476-5926-11-1

    Figure Lengend Snippet: Immunohistochemical staining of liver sections for Abcc3 detection. Frozen livers were cut to 5 μm cryosections and fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS). Sections were blocked in goat serum followed by incubation with anti -Abcc3. Sections were washed with PBS and incubated with goat anti -rat IgG conjugated to Alexfluor 488 (green staining) and rhodamine-conjugated phalloidin (red). Sections were then rinsed with PBS/T, PBS, and water, air dried, and then mounted with Prolong Gold containing DAPI (blue staining). All images are displayed as 200X magnification. It was observed that green staining displaying Abcc3 expression was higher in db/db male and female mice as compared to controls.

    Article Snippet: After incubation with secondary antibody, membranes were washed three times in PBS/T, incubated with ECL + fluorescence Reagent (GE Healthcare, Buckinghamshire, UK), and developed using autoradiography.

    Techniques: Immunohistochemistry, Staining, Incubation, Expressing, Mouse Assay