Quantitative 2-D gel electrophoresis-based expression proteomics of albumin and IgG immunodepleted plasma

Seferovic, Maxim D.; Krughkov, Violet; Pinto, Devanand M.; Han, V. K. M.; Gupta, Madhulika B.

doi:10.1016/j.jchromb.2008.01.052

Cited by 22 publications

(23 citation statements)

References 21 publications

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“…Salting out using ammonium sulfate fractionation (0 -75%) was essential for removal of some interfering proteins and the high salt concentration in AF prior to FFE separation. Rather than conventional albumin depletion (30,31), the salting out procedure proved to be efficient in elimination of albumin to minimize masking effects during FFE separation. Based on the molecular mass marker, although albumin (55-60-kDa band) was still detectable in the concentrated sample (Fig.…”

Section: Assessment Of Igfbp-1 In Ammoniummentioning

confidence: 99%

Functional and Complementary Phosphorylation State Attributes of Human Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) Isoforms Resolved by Free Flow Electrophoresis

Nissum

Shehab²,

Sukop

et al. 2009

Molecular & Cellular Proteomics

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Fetal growth restriction (FGR) is a common disorder in119 . Our data demonstrate that phosphorylation at Ser 119 plays a significant role in modulating affinity of IGFBP-1 for IGF-I. In addition, an altered profile of IGFBP-1 phosphoisoforms was revealed between FGR and healthy pregnancies that may result from potential site-specific phosphorylation. This study provides a strong basis for use of this novel approach in establishing the linkage between phosphorylation of IGFBP-1 and FGR. This overall strategy will also be broadly applicable to other phosphoproteins with clinical and functional significance.

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Section: Assessment Of Igfbp-1 In Ammoniummentioning

confidence: 99%

Functional and Complementary Phosphorylation State Attributes of Human Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) Isoforms Resolved by Free Flow Electrophoresis

Nissum

Shehab²,

Sukop

et al. 2009

Molecular & Cellular Proteomics

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“…Since a large amount of albumin and IgG present in the plasma interferes with separation and identification of low abundant proteins (Seferovic et al, 2008), albumin and IgG were removed by ProteoExtract ™ Albumin/IgG removal kit (cat# 122642, Calbiochem, San Diego, CA) (Bhopale et al, 2011). The proteins were precipitated using ProteoExtract TM protein precipitation kit (cat# 539180, Calbiochem) and measured using RC DC protein assay kit as described earlier.…”

Section: Methodsmentioning

confidence: 99%

Proteomic Profiling of Liver and Plasma in Chronic Ethanol Feeding Model of Hepatic Alcohol Dehydrogenase-Deficient Deer Mice

Bhopale

Amer

Kaphalia

et al. 2017

Alcohol Clin Exp Res

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Background Chronic alcohol abuse, a major risk factor for such diseases as hepatitis and cirrhosis, impairs hepatic alcohol dehydrogenase (ADH, key ethanol metabolizing enzyme). Therefore, differentially altered hepatic and plasma proteomes were identified in chronic ethanol feeding model of hepatic ADH-deficient (ADH−) deer mice to understand the metabolic basis of alcoholic liver disease (ALD). Methods ADH− deer mice were fed 3.5g% ethanol via Lieber-DeCarli liquid diet daily for three months, and histology of the liver assessed. Liver and plasma proteins were separated by 2-dimensional gel electrophoresis. The proteins differentially expressed were identified by MALDI-TOF/TOF mass spectrometry (MS). Results Histology of the liver showed panlobular steatosis and infiltration of T-lymphocytes. Using the criteria of ≥1.5 for fold change (p value ≤0.05) with expectation value (E ≤10−3) and protein score (≥64), eighteen proteins in the livers and five in the plasma of ethanol-fed mice were differentially expressed and identified. Prolyl 4-hydroxylase, cytochrome b-5, endo A cytokeratin, ATP synthase, heat shock 70kD proteins, enoyl CoA hydratase, stress-70 protein, peroxiredoxin 1 and ornithine carbamoyl-transferase were up regulated in the livers. However, carbonic anhydrase 3, mitochondrial ATP synthase, aldolase 2, actin γ, laminin receptor and carbamoyl-phosphate synthase were down regulated. Contrary to the increased expression of creatine kinase m-type, a decreased expression of serine protease inhibitor A3A precursor, sulfated glycoprotein-2 (clusterin) and apolipoprotein E isoforms were found in the plasma of ethanol group. Conclusions Chronic ethanol feeding in ADH− deer mice causes steatosis and infiltration of T-lymphocytes in the livers along with increased expression of proteins involved in ER stress, fibrosis, fatty acid β oxidation and biogenesis, and decreased expression of proteins involved in ATP synthesis, carbohydrate metabolism, in cell regulation and architecture. Reduced expression of various carrier proteins as found in the plasma of ethanol group has a biomarker potential.

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“…First-dimension separation methods include isoelectric focusing electrophoresis (IEF) [1][2][3][4][5], on-line multiple-junction capillary isoelectric focusing fractionator (OMJ-CIEF) [6], one-dimensional and two-dimensional gel electrophoresis [7][8][9], IEF with superficially porous liquid chromatography (IEF-SPLC) [10], off-line and on-line high performance liquid chromatographic (HPLC) separation [11][12][13], two-dimensional (2D)-LC systems with online fractionation of proteins into a series of small trapping columns [14], 2D-LC-UV/MS analysis [15] and three-dimensional approaches [16,17]. All these methods provide important tools for identifying and quantifying proteomic difference between multiple biological conditions.…”

Section: Introductionmentioning

confidence: 99%

An off-line high pH reversed-phase fractionation and nano-liquid chromatography–mass spectrometry method for global proteomic profiling of cell lines

Wang

Sun

Zhang

et al. 2015

Journal of Chromatography B

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Quantitative 2-D gel electrophoresis-based expression proteomics of albumin and IgG immunodepleted plasma

Cited by 22 publications

References 21 publications

Functional and Complementary Phosphorylation State Attributes of Human Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) Isoforms Resolved by Free Flow Electrophoresis

Functional and Complementary Phosphorylation State Attributes of Human Insulin-like Growth Factor-Binding Protein-1 (IGFBP-1) Isoforms Resolved by Free Flow Electrophoresis

Proteomic Profiling of Liver and Plasma in Chronic Ethanol Feeding Model of Hepatic Alcohol Dehydrogenase-Deficient Deer Mice

An off-line high pH reversed-phase fractionation and nano-liquid chromatography–mass spectrometry method for global proteomic profiling of cell lines

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