pbs solution  (Millipore)

 
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    Name:
    Phosphate buffer solution
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    Catalog Number:
    1805
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    Structured Review

    Millipore pbs solution

    https://www.bioz.com/result/pbs solution/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Related Articles

    Flow Cytometry:

    Article Title: Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages
    Article Snippet: Supernatants were collected and pooled (n = 4) for aortic ring assay stimulation. .. Flow cytometry One million mouse PECs were incubated in ice-cold PBS medium (Blocking buffer) containing 5 mM EDTA (Sigma Aldrich, France), 1 % FCS, 3 % normal rat and mouse serum, and 10 % mouse Seroblock (anti CD16/CD32, Abd Serotec, UK). .. Macrophages were stained 25 min on ice with APC-conjugated rat anti-mouse F4/80 (MCA497APC) and PE-coupled rat anti-mouse CD11b (MCA711PE, Serotec).

    Cytometry:

    Article Title: Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages
    Article Snippet: Supernatants were collected and pooled (n = 4) for aortic ring assay stimulation. .. Flow cytometry One million mouse PECs were incubated in ice-cold PBS medium (Blocking buffer) containing 5 mM EDTA (Sigma Aldrich, France), 1 % FCS, 3 % normal rat and mouse serum, and 10 % mouse Seroblock (anti CD16/CD32, Abd Serotec, UK). .. Macrophages were stained 25 min on ice with APC-conjugated rat anti-mouse F4/80 (MCA497APC) and PE-coupled rat anti-mouse CD11b (MCA711PE, Serotec).

    Incubation:

    Article Title: Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages
    Article Snippet: Supernatants were collected and pooled (n = 4) for aortic ring assay stimulation. .. Flow cytometry One million mouse PECs were incubated in ice-cold PBS medium (Blocking buffer) containing 5 mM EDTA (Sigma Aldrich, France), 1 % FCS, 3 % normal rat and mouse serum, and 10 % mouse Seroblock (anti CD16/CD32, Abd Serotec, UK). .. Macrophages were stained 25 min on ice with APC-conjugated rat anti-mouse F4/80 (MCA497APC) and PE-coupled rat anti-mouse CD11b (MCA711PE, Serotec).

    Article Title: Absence of Intestinal PPAR? Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2-Dependent Pathway
    Article Snippet: The supernatant was collected for use as PBS extracts for the analysis of secreted proteins, while the pellet was re-extracted in lysis buffer (10 mM Tris-HCl [pH 8], 150 mM NaCl, 1% Triton X-100, and protease inhibitors) to obtain Triton X-100 extracts . .. For partial purification of MMP-9 and MMP-2, PBS extracts of the respective samples were incubated with gelatin-agarose beads (Sigma) at 4°C for 1 h followed by centrifugation at 1500 rpm. ..

    Blocking Assay:

    Article Title: Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages
    Article Snippet: Supernatants were collected and pooled (n = 4) for aortic ring assay stimulation. .. Flow cytometry One million mouse PECs were incubated in ice-cold PBS medium (Blocking buffer) containing 5 mM EDTA (Sigma Aldrich, France), 1 % FCS, 3 % normal rat and mouse serum, and 10 % mouse Seroblock (anti CD16/CD32, Abd Serotec, UK). .. Macrophages were stained 25 min on ice with APC-conjugated rat anti-mouse F4/80 (MCA497APC) and PE-coupled rat anti-mouse CD11b (MCA711PE, Serotec).

    Infection:

    Article Title: The Recombinant Form of Trypanosoma cruzi P21 Controls Infection by Modulating Host Immune Response
    Article Snippet: Reactions were processed in ABI7300 equipment (Applied Biosystems) under the following conditions: 50°C for 2 min, 90°C for 10 min, 40 cycles at 95°C for 15 s and 60°C for 45 s and 72°C for 30 s. After the final elongation of qPCR, samples were submitted to temperature variation from 50 to 95°C, with a gradual increase of 0.5°C/s to obtain the melting temperature (Tm) and non-specific products. .. Cytokine Immunoassays For evaluation of cytokine levels in cardiac and spleen tissues harvested from control and T. cruzi infected mice, tissue sections were immersed in PBS solution containing protease inhibitor cocktail (1 tablet diluted in 50 ml of PBS-Complete, Sigma Aldrich). .. After homogenization with tissue homogenizer, the quantification of cytokines and total proteins was performed.

    Mouse Assay:

    Article Title: The Recombinant Form of Trypanosoma cruzi P21 Controls Infection by Modulating Host Immune Response
    Article Snippet: Reactions were processed in ABI7300 equipment (Applied Biosystems) under the following conditions: 50°C for 2 min, 90°C for 10 min, 40 cycles at 95°C for 15 s and 60°C for 45 s and 72°C for 30 s. After the final elongation of qPCR, samples were submitted to temperature variation from 50 to 95°C, with a gradual increase of 0.5°C/s to obtain the melting temperature (Tm) and non-specific products. .. Cytokine Immunoassays For evaluation of cytokine levels in cardiac and spleen tissues harvested from control and T. cruzi infected mice, tissue sections were immersed in PBS solution containing protease inhibitor cocktail (1 tablet diluted in 50 ml of PBS-Complete, Sigma Aldrich). .. After homogenization with tissue homogenizer, the quantification of cytokines and total proteins was performed.

    Article Title: STAT4 activation by leukemia inhibitory factor confers a therapeutic effect on intestinal inflammation
    Article Snippet: The LIF concentration in the supernatant was quantified by an ELISA kit (R & D Systems, UK) and normalized to the explant weight. .. Colon tissues dissected from control or DSS‐challenged mice were washed in PBS buffer and were then cut into pieces and digested at 37°C for 1 h in DMEM supplemented with 1% FBS, penicillin (100 U/ml), streptomycin (100 U/ml), collagenase type XI (0.2 mg/ml, Sigma), and dispase II (200 μg/ml, Roche). .. After digestion, the crypts containing IECs were isolated from the supernatant of the digestion buffer by centrifugation at 300 × g for 5 min.

    Protease Inhibitor:

    Article Title: The Recombinant Form of Trypanosoma cruzi P21 Controls Infection by Modulating Host Immune Response
    Article Snippet: Reactions were processed in ABI7300 equipment (Applied Biosystems) under the following conditions: 50°C for 2 min, 90°C for 10 min, 40 cycles at 95°C for 15 s and 60°C for 45 s and 72°C for 30 s. After the final elongation of qPCR, samples were submitted to temperature variation from 50 to 95°C, with a gradual increase of 0.5°C/s to obtain the melting temperature (Tm) and non-specific products. .. Cytokine Immunoassays For evaluation of cytokine levels in cardiac and spleen tissues harvested from control and T. cruzi infected mice, tissue sections were immersed in PBS solution containing protease inhibitor cocktail (1 tablet diluted in 50 ml of PBS-Complete, Sigma Aldrich). .. After homogenization with tissue homogenizer, the quantification of cytokines and total proteins was performed.

    Injection:

    Article Title: Laminin-111 improves muscle repair in a mouse model of merosin-deficient congenital muscular dystrophy
    Article Snippet: The muscles were harvested either at 0, 4, 10 or 28 days after CTX injection for analysis. .. TA muscles were damaged at Day 0, three days after laminin or PBS treatments by i.m. injection of 100 µL of a 10 µmol/L CTX solution (C3987; Sigma, St. Louis, MO, USA) in PBS. .. At 4, 10, or 28 days after CTX-induced injury, the mice were euthanized and TA muscles harvested for analysis.

    Staining:

    Article Title: CD34+ Cells Represent Highly Functional Endothelial Progenitor Cells in Murine Bone Marrow
    Article Snippet: After washing three times with PBS for 5 min each, cells were incubated with secondary antibodies prepared at 1∶500 in antibody dilution buffer: Alexa 488 donkey anti-goat IgG, Alexa 488 goat anti-rabbit IgG and Alexa 488 goat anti-rat IgG (MP/Invitrogen) for 30 min at RT. .. After secondary antibodies were removed and cells were washed with PBS for three times, DAPI solution (Sigma, 1∶5000) was added and nuclei were stained for 10 min at RT. .. Mounting medium and a cover-slip were added to the glass slide followed by sealing samples with nail varnish before evaluation of the staining results under fluorescence microscopy.

    Purification:

    Article Title: Absence of Intestinal PPAR? Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2-Dependent Pathway
    Article Snippet: The supernatant was collected for use as PBS extracts for the analysis of secreted proteins, while the pellet was re-extracted in lysis buffer (10 mM Tris-HCl [pH 8], 150 mM NaCl, 1% Triton X-100, and protease inhibitors) to obtain Triton X-100 extracts . .. For partial purification of MMP-9 and MMP-2, PBS extracts of the respective samples were incubated with gelatin-agarose beads (Sigma) at 4°C for 1 h followed by centrifugation at 1500 rpm. ..

    Centrifugation:

    Article Title: Absence of Intestinal PPAR? Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2-Dependent Pathway
    Article Snippet: The supernatant was collected for use as PBS extracts for the analysis of secreted proteins, while the pellet was re-extracted in lysis buffer (10 mM Tris-HCl [pH 8], 150 mM NaCl, 1% Triton X-100, and protease inhibitors) to obtain Triton X-100 extracts . .. For partial purification of MMP-9 and MMP-2, PBS extracts of the respective samples were incubated with gelatin-agarose beads (Sigma) at 4°C for 1 h followed by centrifugation at 1500 rpm. ..

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  • 97
    Millipore pbs medium
    sDLL4 activates Notch signaling and induces proangiogenic mediators in macrophages in vitro: flow <t>cytometry</t> of F4/80 + CD11b + macrophages ( a ), Notch1 ( b ), Notch4 ( c ) and DLL4 ( d ). HES-1 mRNA expression by PECs cultivated on sDLL4 coated plates (DLL4an = DLL4anchored) ( e , <t>PBS,</t> white , n = 3, DLL4an, black , n = 3). HES-1 mRNA expression of soluble (s) DLL4 stimulated macrophages ( f , PBS, white , n = 6 and sDLL4, black , n = 4). Western blotting of Notch-4 intracellular domain (N4-ICD) in macrophages stimulated with PBS or sDLL4 ( g ). mRNA expression in sDLL4 and sDLL4 and DAPT treated C57BL/6j activated macrophages of HES-1 ( h ), VEGF ( i ), IL-1β ( j ), IL-6 ( k ) and TNF-α ( l ). (LPS n = 4 white , LPS + sDLL4 n = 4 black , LPS + sDLL4 +DAPT n = 4 grey ) * p
    Pbs Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs medium/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    97
    Millipore phosphate buffered saline pbs solution
    Comparison of permeabilization buffer and paraformaldehyde fixation on minimum inhibition response (solid squares), maximum inhibition response (solid triangles), and Z' factor (red circles) of Plasmodium falciparum Dd2 strain parasites at varying parasitemia and 0.3% hematocrit. Combinations of <t>Tris-HCl</t> buffer with fixation ( A ), Tris-HCl buffer without fixation ( B ), phosphate-buffered saline <t>(PBS)</t> buffer with fixation ( C ), and PBS buffer without fixation ( D ) are shown. Where a linear relationship between parasitemia and minimum inhibition response was observed, linear regression (dotted) line and r 2 values were included.
    Phosphate Buffered Saline Pbs Solution, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phosphate buffered saline pbs solution/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    97
    Millipore pbs edta
    Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as <t>PBS-EDTA,</t> to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.
    Pbs Edta, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbs edta/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbs edta - by Bioz Stars, 2021-04
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    Image Search Results


    sDLL4 activates Notch signaling and induces proangiogenic mediators in macrophages in vitro: flow cytometry of F4/80 + CD11b + macrophages ( a ), Notch1 ( b ), Notch4 ( c ) and DLL4 ( d ). HES-1 mRNA expression by PECs cultivated on sDLL4 coated plates (DLL4an = DLL4anchored) ( e , PBS, white , n = 3, DLL4an, black , n = 3). HES-1 mRNA expression of soluble (s) DLL4 stimulated macrophages ( f , PBS, white , n = 6 and sDLL4, black , n = 4). Western blotting of Notch-4 intracellular domain (N4-ICD) in macrophages stimulated with PBS or sDLL4 ( g ). mRNA expression in sDLL4 and sDLL4 and DAPT treated C57BL/6j activated macrophages of HES-1 ( h ), VEGF ( i ), IL-1β ( j ), IL-6 ( k ) and TNF-α ( l ). (LPS n = 4 white , LPS + sDLL4 n = 4 black , LPS + sDLL4 +DAPT n = 4 grey ) * p

    Journal: Angiogenesis

    Article Title: Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages

    doi: 10.1007/s10456-012-9290-0

    Figure Lengend Snippet: sDLL4 activates Notch signaling and induces proangiogenic mediators in macrophages in vitro: flow cytometry of F4/80 + CD11b + macrophages ( a ), Notch1 ( b ), Notch4 ( c ) and DLL4 ( d ). HES-1 mRNA expression by PECs cultivated on sDLL4 coated plates (DLL4an = DLL4anchored) ( e , PBS, white , n = 3, DLL4an, black , n = 3). HES-1 mRNA expression of soluble (s) DLL4 stimulated macrophages ( f , PBS, white , n = 6 and sDLL4, black , n = 4). Western blotting of Notch-4 intracellular domain (N4-ICD) in macrophages stimulated with PBS or sDLL4 ( g ). mRNA expression in sDLL4 and sDLL4 and DAPT treated C57BL/6j activated macrophages of HES-1 ( h ), VEGF ( i ), IL-1β ( j ), IL-6 ( k ) and TNF-α ( l ). (LPS n = 4 white , LPS + sDLL4 n = 4 black , LPS + sDLL4 +DAPT n = 4 grey ) * p

    Article Snippet: Flow cytometry One million mouse PECs were incubated in ice-cold PBS medium (Blocking buffer) containing 5 mM EDTA (Sigma Aldrich, France), 1 % FCS, 3 % normal rat and mouse serum, and 10 % mouse Seroblock (anti CD16/CD32, Abd Serotec, UK).

    Techniques: In Vitro, Flow Cytometry, Cytometry, Expressing, Western Blot

    Dissolution tests of MSG in PBS at pH 7.4 and 37 °C.

    Journal: Pharmaceutics

    Article Title: PCL/Mesoglycan Devices Obtained by Supercritical Foaming and Impregnation

    doi: 10.3390/pharmaceutics11120631

    Figure Lengend Snippet: Dissolution tests of MSG in PBS at pH 7.4 and 37 °C.

    Article Snippet: Additionally, PBS buffer in which MSG has been dissolved from PCL films was filtered by sterile syringe 0.22 µm filters (Sigma Aldrich, Milan, Italy) and administered to cells.

    Techniques:

    Comparison of permeabilization buffer and paraformaldehyde fixation on minimum inhibition response (solid squares), maximum inhibition response (solid triangles), and Z' factor (red circles) of Plasmodium falciparum Dd2 strain parasites at varying parasitemia and 0.3% hematocrit. Combinations of Tris-HCl buffer with fixation ( A ), Tris-HCl buffer without fixation ( B ), phosphate-buffered saline (PBS) buffer with fixation ( C ), and PBS buffer without fixation ( D ) are shown. Where a linear relationship between parasitemia and minimum inhibition response was observed, linear regression (dotted) line and r 2 values were included.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Development and Optimization of a Novel 384-Well Anti-Malarial Imaging Assay Validated for High-Throughput Screening

    doi: 10.4269/ajtmh.2012.11-0302

    Figure Lengend Snippet: Comparison of permeabilization buffer and paraformaldehyde fixation on minimum inhibition response (solid squares), maximum inhibition response (solid triangles), and Z' factor (red circles) of Plasmodium falciparum Dd2 strain parasites at varying parasitemia and 0.3% hematocrit. Combinations of Tris-HCl buffer with fixation ( A ), Tris-HCl buffer without fixation ( B ), phosphate-buffered saline (PBS) buffer with fixation ( C ), and PBS buffer without fixation ( D ) are shown. Where a linear relationship between parasitemia and minimum inhibition response was observed, linear regression (dotted) line and r 2 values were included.

    Article Snippet: Hypoxanthine, 1 M NaOH, D-sorbitol, chloroquine diphosphate salt, artemisinin, 10× phosphate buffered saline (PBS) solution, EDTA, 0.5 M solution, Tris-HCl, saponin (from Quillaja Bark), Triton X-100, Giemsa stain, paraformaldehyde, dimethylformamide, and Petri dishes were purchased from Sigma Aldrich (St. Louis, MO).

    Techniques: Inhibition

    Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as PBS-EDTA, to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: Representative absorbance spectra of AMB-PEG 2 and unconjugated AMB formulations prepared in buffers with varying hydrophobicity. 20 mM AMB-PEG 2 and unconjugated AMB formulations were prepared in DMSO and resuspended in 20% and 48% ACN buffers containing 4.3% acetic acid, as well as PBS-EDTA, to a final concentration of 2 mM AMB. As buffer hydrophobicity increases with higher ACN concentrations, the A 348 /A 409 ratio decreases, implying that AMB-PEG is increasingly in its monomeric form. As AMB-PEG 1 and 2 have similar UV-visible absorption profiles, with identical peak height ratios in all buffers tested, data for AMB-PEG 1 is not shown. AMB-PEG formulations that have been subjected to buffer exchange to PBS-EDTA through a 10 kDa centrifugal filter have the same UV-visible absorption spectra as the initial formulation of AMB-PEG in PBS-EDTA, which contains 10% DMSO.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Concentration Assay, Buffer Exchange

    (A) Reverse phase chromatogram of AMB-PEG and unconjugated AMB, with eluted peaks detected at 406 nm. 50 μL of AMB-PEG 1 and 2, and 10 μL of unconjugated AMB dispersed in PBS-EDTA and 48% ACN respectively were injected into a C18 reverse phase column and eluted isocratically in a 48% ACN buffer at a flow rate of 0.5 ml/min for 40 minutes. Peaks were detected at 406 nm. The AMB-PEG conjugate has a shorter retention time, implying that it is more hydrophilic. From the AMB-PEG samples, AMB-PEG conjugate and free AMB (based on the retention time of unconjugated AMB) fractions were collected for further analysis via size exclusion chromatography. (B) Size exclusion chromatogram of AMB-PEG 2 and unconjugated AMB, as well as the relevant fractions collected from RPC, in a 20% ACN mobile phase. 20 mM of AMB-PEG 2 formulations and unconjugated AMB in DMSO were prepared and resuspended at 2 mM in a 20% ACN buffer. 50 μL of these samples (unconjugated AMB diluted tenfold prior to analysis) and the peak fractions collected previously from RPC were passed through a size exclusion column and eluted peaks were detected at 406 nm. Unconjugated AMB was eluted later compared to the AMB-PEG conjugate, implying that it has a smaller hydrodynamic volume compared to AMB-PEG in 20% ACN. The previously collected AMB-PEG conjugate and free AMB peak fractions had similar retention times as the AMB-PEG 2 formulation and unconjugated AMB samples respectively, thus verifying their respective peak identities. AMB-PEG 1 and 2 have identical elution profiles. (C) Size exclusion chromatogram of AMB-PEG and unconjugated AMB dispersed in PBS-EDTA. 3 μL of 2 mM AMB-PEG formulations that have been retained by 10 kDa centrifugal filters (Millipore) and 50 μL of the supernatant obtained from unconjugated AMB were analysed using a Superdex 75 size exclusion column and eluted peaks detected at 406 nm. In a PBS-EDTA mobile phase, unconjugated AMB has a shorter retention time of 20 minutes compared to AMB-PEG at 40 minutes, implying that AMB-PEG has a smaller hydrodynamic volume under these experimental conditions.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: (A) Reverse phase chromatogram of AMB-PEG and unconjugated AMB, with eluted peaks detected at 406 nm. 50 μL of AMB-PEG 1 and 2, and 10 μL of unconjugated AMB dispersed in PBS-EDTA and 48% ACN respectively were injected into a C18 reverse phase column and eluted isocratically in a 48% ACN buffer at a flow rate of 0.5 ml/min for 40 minutes. Peaks were detected at 406 nm. The AMB-PEG conjugate has a shorter retention time, implying that it is more hydrophilic. From the AMB-PEG samples, AMB-PEG conjugate and free AMB (based on the retention time of unconjugated AMB) fractions were collected for further analysis via size exclusion chromatography. (B) Size exclusion chromatogram of AMB-PEG 2 and unconjugated AMB, as well as the relevant fractions collected from RPC, in a 20% ACN mobile phase. 20 mM of AMB-PEG 2 formulations and unconjugated AMB in DMSO were prepared and resuspended at 2 mM in a 20% ACN buffer. 50 μL of these samples (unconjugated AMB diluted tenfold prior to analysis) and the peak fractions collected previously from RPC were passed through a size exclusion column and eluted peaks were detected at 406 nm. Unconjugated AMB was eluted later compared to the AMB-PEG conjugate, implying that it has a smaller hydrodynamic volume compared to AMB-PEG in 20% ACN. The previously collected AMB-PEG conjugate and free AMB peak fractions had similar retention times as the AMB-PEG 2 formulation and unconjugated AMB samples respectively, thus verifying their respective peak identities. AMB-PEG 1 and 2 have identical elution profiles. (C) Size exclusion chromatogram of AMB-PEG and unconjugated AMB dispersed in PBS-EDTA. 3 μL of 2 mM AMB-PEG formulations that have been retained by 10 kDa centrifugal filters (Millipore) and 50 μL of the supernatant obtained from unconjugated AMB were analysed using a Superdex 75 size exclusion column and eluted peaks detected at 406 nm. In a PBS-EDTA mobile phase, unconjugated AMB has a shorter retention time of 20 minutes compared to AMB-PEG at 40 minutes, implying that AMB-PEG has a smaller hydrodynamic volume under these experimental conditions.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Injection, Flow Cytometry, Size-exclusion Chromatography

    (A) AMB-PEG reaction scheme. The NHS ester on MS(PEG) 4 reacts with the primary amine (–NH2) group on AMB at a 1:1 molar ratio, generating conjugated AMB-PEG via amide bond formation. (B) Solubility of AMB-PEG at varying AMB:PEG molar ratios. AMB-PEG mixtures and unreacted AMB were prepared in DMSO, incubated for 2 hours and then (i) dispersed to a final concentration of 2 mM in PBS-EDTA, containing 10% DMSO by volume. (ii) The mixtures were then centrifuged to facilitate the observation of insoluble precipitates. Higher molar ratios yielded a suspension of yellow particles that precipitated upon centrifugation, similar to what was observed with unconjugated AMB.

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: (A) AMB-PEG reaction scheme. The NHS ester on MS(PEG) 4 reacts with the primary amine (–NH2) group on AMB at a 1:1 molar ratio, generating conjugated AMB-PEG via amide bond formation. (B) Solubility of AMB-PEG at varying AMB:PEG molar ratios. AMB-PEG mixtures and unreacted AMB were prepared in DMSO, incubated for 2 hours and then (i) dispersed to a final concentration of 2 mM in PBS-EDTA, containing 10% DMSO by volume. (ii) The mixtures were then centrifuged to facilitate the observation of insoluble precipitates. Higher molar ratios yielded a suspension of yellow particles that precipitated upon centrifugation, similar to what was observed with unconjugated AMB.

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Mass Spectrometry, Solubility, Incubation, Concentration Assay, Centrifugation

    Aqueous solubility of AMB-PEG 1 and 2. Increasing amounts of AMB-PEG formulations were added to PBS-EDTA, and post-centrifugation, the concentration of soluble AMB-PEG in the supernatants quantified based on their absorbance at 365 nm. Error bars denote the standard deviation between 3 independent formulations. Dotted line represents the theoretical condition where AMB formulation is completely soluble (i.e. concentration of total AMB = concentration of soluble AMB).

    Journal: PLoS ONE

    Article Title: Characterization of a Polyethylene Glycol-Amphotericin B Conjugate Loaded with Free AMB for Improved Antifungal Efficacy

    doi: 10.1371/journal.pone.0152112

    Figure Lengend Snippet: Aqueous solubility of AMB-PEG 1 and 2. Increasing amounts of AMB-PEG formulations were added to PBS-EDTA, and post-centrifugation, the concentration of soluble AMB-PEG in the supernatants quantified based on their absorbance at 365 nm. Error bars denote the standard deviation between 3 independent formulations. Dotted line represents the theoretical condition where AMB formulation is completely soluble (i.e. concentration of total AMB = concentration of soluble AMB).

    Article Snippet: AMB-PEG Particle Size Determination Using DLS 2 mM solutions of AMB-PEG were prepared in PBS-EDTA, and diafiltration carried out with PBS-EDTA using 10 kDa centrifugal filters (Millipore).

    Techniques: Solubility, Centrifugation, Concentration Assay, Standard Deviation