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Heparin-sepharose

Heparin-sepharose is a chromatographic material commonly used in the purification and analysis of biomolecules, particularly proteins.
It consists of heparin, a sulfated glycosaminoglycan, immobilized on a sepharose matrix.
Heparin-sepharose exploits the affinity of heparin for various proteins, allowing for the selective capture and separation of these molecules based on their interactions with heparin.
This technique is widely employed in the study of growth factors, enzymes, and other proteins of biological importance.
Heparin-sepharose chromatography is a versatile and effective tool for researchers investigating protein structure, function, and purification.

Most cited protocols related to «Heparin-sepharose»

Purification of Retroviral Integrase Proteins

Cited 17 times

Endonuclease A-deficient Escherichia coli strain PC2 [BL21(DE3), endA::Tet^R, T1^R, pLysS], recovered as a spontaneous T1 phage-resistant mutant of PC1 (51 (link),52 (link)), was used for protein production. Shake flask cultures of PC2 transformed with various expression constructs were grown in LB medium at 30°C to an A₆₀₀ of 0.9–1.0 prior to induction with 0.25 mM isopropyl-thio-β-d-galactopyranoside. Following 4 h induction at 25–28°C, bacteria were harvested and stored at −70°C.
To isolate His₆-tagged retroviral IN proteins, thawed bacterial paste was sonicated in buffer B (1 M NaCl, 7.5 mM CHAPS, 50 mM Tris–HCl, pH 7.4) containing 0.5 mM phenylmethlysulfonyl fluoride and 15 mM imidazole. Crude extracts pre-cleared by centrifugation were incubated with Ni NTA agarose (Qiagen). The resin was extensively washed in buffer B containing 15 mM imidazole, and His₆-tagged proteins were eluted with 200 mM imidazole in buffer B. IN-containing fractions diluted with 3 volumes of 50 mM Tris–HCl, pH 7.4 were injected into a 5 ml HiTrap Heparin column (GE Healthcare), and bound proteins were eluted with a linear gradient of 0.25–1 M NaCl in 50 mM Tris–HCl, pH 7.4. Immediately after elution, 10 mM DTT was added to each fraction and NaCl concentration was adjusted to 1 M. HIV-1, BIV and EIAV INs produced to carry cleavable N-terminal His₆-tags were digested with thrombin (3 NIH units of thrombin per mg of His₆-tagged IN) for 3 h at 25°C in the presence of 20 mM β-mercaptoethanol or HRV14 3C protease (20 μg of protease per mg of His₆-tagged IN) for 6–12 h at 4°C in the presence of 20 mM DTT; the tag-free INs were then purified by chromatography on Heparin sepharose as above.
Wild type and D366N LEDGF were made as previously described (24 (link),25 (link),31 (link)). Purified proteins supplemented with 10% glycerol were flash-frozen in liquid nitrogen and stored at −70°C. Protein concentration was determined using Bradford assay (Bio-Rad Laboratories) with a BSA standard. Where indicated, molar concentrations refer to monomer protein forms.

, & Cherepanov P. (2006). LEDGF/p75 interacts with divergent lentiviral integrases and modulates their enzymatic activity in vitro. Nucleic Acids Research, 35(1), 113-124.

Publication 2006

2-Mercaptoethanol 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate Bacteria Bacteriophages Biological Assay Buffers Centrifugation Chromatography Complex Extracts Endonuclease Escherichia coli Fluorides Freezing Galactose Glycerin Heparin heparin-sepharose his6 tag HIV-1 imidazole Infectious Anemia Virus, Equine lens epithelium-derived growth factor Molar Nitrogen Paste Peptide Hydrolases Proteins Resins, Plant Retroviridae Proteins Sepharose Sodium Chloride Strains Thrombin Tremor Tromethamine

Thioglycollate-Induced Peritoneal Macrophage Activation

Cited 11 times

To activate peritoneal macrophages, mice were administered 1.5 ml of 4% thioglycollate broth intraperitoneally. After 72 h, cells were harvested by peritoneal lavage with 5 ml cold HBSS with 10 U/ml heparin, and plated in RPMI 1640 medium supplemented with 10% bovine calf serum. After 2 h, nonadherent cells were removed by washing and adherent cells were stimulated with 10 μg/ml LPS (Sigma Chemical Co., St. Louis, MO) for 4 h. Cells were radiolabeled with 2 mCi/ml [³⁵S]methionine (NEN-DuPont, Boston, MA) in the presence of 10 μg/ml LPS for an additional 4 h. Medium was collected, diluted with an equal volume of RIPA buffer, and precleared with normal rabbit serum and protein A–Sepharose beads (Bio-Rad Laboratories, Hercules, CA). Immune precipitations were then performed sequentially with anti–murine MCP-1 (21 (link)) and anti-KC (27 (link)). Anti-MCP-3 (28 (link)) (a gift from R. Bravo, Bristol-Myers Squibb, Princeton, NJ) was used in a separate experiment. Similar results were obtained without thioglycollate treatment, although the levels of chemokine synthesis were much lower in wild-type and MCP-1^−/− mice, suggesting that thioglycollate priming is necessary for a full response to LPS in resident peritoneal macrophages.

Lu B., Rutledge B.J., Gu L., Fiorillo J., Lukacs N.W., Kunkel S.L., North R., Gerard C, & Rollins B.J. (1998). Abnormalities in Monocyte Recruitment and Cytokine Expression in Monocyte Chemoattractant Protein 1–deficient Mice. The Journal of Experimental Medicine, 187(4), 601-608.

Publication 1998

Anabolism Bos taurus Buffers CCL2 protein, human CCL7 protein, human Cells Chemokine Cold Temperature Hemoglobin, Sickle Heparin Immunoprecipitation Macrophages, Peritoneal Methionine Mus Peritoneal Lavage Rabbits Radioimmunoprecipitation Assay Serum Staphylococcal protein A-sepharose Thioglycolates

Borrelia Immune Evasion Mechanisms

Cited 11 times

Bacterial Strains and Culture Condition—B. garinii strain G1 was originally isolated from the cerebrospinal fluid of a Lyme
disease patient and is highly susceptible to complement-mediated killing
in vitro (7 (link)).
Borrelia were grown at 33 °C for 5–6 days up to cell
densities of 1 × 10⁷ ml^-1 in modified
Barbour-Stoenner-Kelly (BSK) medium as described previously
(19 (link)). Escherichia
coli JM109 was used as host for cloning and purification of recombinant
proteins.
Site-directed Mutagenesis, Sequence Analysis, and Generation of
Recombinant Proteins—To introduce single or double amino acid
substitutions into the BbCRASP-2-encoding cspZ gene, a site-directed
mutagenesis approach was conducted as described by the QuikChange™
protocol (Stratagene). The mutagenic high-performance liquid
chromatography-purified primers employed in this study were purchased from
Sigma-Aldrich (Steinheim, Germany) and listed in supplemental Table S1.
Briefly, PCR reactions were carried out for 18 cycles (95 °C for 30 s, 55
°C for 30 s, and 68 °C for 13 min) using 100 ng of expression vector
pGEX CSPZ, 125 ng of each mutagenic primer, and 2.5 units of Pfu DNA
polymerase (Stratagene) in a final volume of 50 μl. For the introduction of
site-specific mutations into shuttle vector pCSPZ (previously referred as
pCRASP-2 (31 (link))), thermal
cycling was performed for 18 cycles at 95 °C for 30 s, 55 °C for 60 s,
and 68 °C for 16 min. Before transformation of E. coli, the
reactions were treated with 10 units of DpnI (New England Biolabs, Frankfurt,
Germany) for 1 h at 37 °C. All mutations introduced into the cspZ gene were verified by DNA sequencing of both DNA strands. Proteins were
purified using glutathione-Sepharose columns, with conditions as recommended
by the manufacturer (GE Healthcare).
Binding and Inhibition Assay—Purified GST-BbCRASP-2, mutated
GST-BbCRASP-2 proteins, and GST (2.5 ng/μl each) were immobilized overnight
at 4 °C using Immobilizier™ glutathione microtiter plates (Nunc,
Wiesbaden, Germany). Non-specific binding sites were blocked with 0.2% gelatin
in PBS for 6 h at 4 °C. CFH (Calbiochem, Darmstadt, Germany) or purified
CFHL1 (16 (link)) (5 μg/ml each)
was added to the wells and left overnight at 4 °C. After addition of
polyclonal anti-CFH antibodies (Calbiochem) for 2 h at room temperature,
protein complexes were identified using a secondary peroxidase-conjugated
anti-goat IgG antibody. Reactions were developed with 1,2-phenylenediamine
dihydrochloride (Sigma-Aldrich).
Influences of heparin and salt on CFH binding were analyzed by ELISA.
Purified GST-BbCRASP-2 (15 ng/μl) was immobilized onto wells of a
microtiter plate (MaxiSorb, Nunc) overnight at 4 °C. Following three
washing steps with PBS containing 0.05% Tween 20 (PBST), nonspecific binding
sites were blocked with 0.2% gelatin in PBS for 6 h at 4 °C. Plates
covered with BbCRASP-2 fusion protein were incubated overnight at 4 °C
with CFH (0.25 ng/well) in PBST containing increasing concentrations of either
low molecular weight heparin (Sigma-Aldrich, 0.5–16 μg/ml) or NaCl
(29 mm to 1.16 m). After washing three times in PBS,
binding of CFH was detected using mAb VIG8 antibody
(35 ). The experiments were
conducted at least three times, and the means ± S.D. were
calculated.
SDS-PAGE, Ligand Affinity Blot, and Western Blot
Analysis—Borrelial cell extracts (15 μg) were subjected to 10%
Tris/Tricine-SDS-PAGE under reducing conditions and transferred to
nitrocellulose. Binding of CFH and CFHL1 to borrelial proteins was assessed by
ligand affinity blotting as previously described
(19 (link)).
For Western blot analysis, membranes were incubated for 60 min at room
temperature with either mAb L41 1C11 (FlaB)
(36 (link)) or polyclonal anti-GST
antibody (GE Healthcare, Germany) as described elsewhere
(19 (link)).
Transformation of Serum-sensitive B. garinii and Characterization of
Transformants—High passage, non-infectious B. garinii strain G1 was grown in 100 ml of BSK medium and harvested at mid-exponential
phase (5 × 10⁷ to 1 × 10⁸ cells/ml).
Electrocompetent cells were prepared as described previously
(37 ) with slight
modifications. Briefly, 50-μl aliquots of competent B. garinii strain G1 cells were electroporated at 12.5 kV/cm in 2-mm cuvettes with 10
μg of plasmid DNA. For control purpose B. garinii strain G1 cells
also were transformed with pKFSS1 vector
(38 (link)) alone. Cells were
immediately diluted into 10 ml of BSK medium and incubated with antibiotic
selection at 33 °C for 48–72 h. Bacteria were then diluted into 100
ml of BSK medium containing streptomycin (25 μg/ml), and 200-μl aliquots
were plated into 96-well cell culture plates (Corning) for selection of
transformants. Several clones selected were expanded in 1 ml of fresh BSK
medium without antibiotic selection for 7 days, and then transferred into 10
ml of fresh BSK medium containing streptomycin (50 μg/ml).
The cspZ genes of transformed B. garinii G1 strains were
detected by PCR with specific primers (Table S1). Spirochetes (100 μl) were
sedimented by centrifugation, washed with PBS, and suspended in 50 μl of
water. Five microliters of suspension was amplified by PCR using
oligonucleotide primers at final concentrations of 100 nm each,
plus 200 μm dNTPs. PCR was carried out for 25 cycles using the
following parameters: denaturation at 94 °C for 1 min, annealing at 50
°C for 1 min, and extension at 72 °C for 1 min.
Plasmid DNA was prepared from the presumptive E. coli transformants with the Wizard® Plus SV Miniprep DNA purification system
(Promega, Mannheim, Germany), and DNA inserts were sequenced by a commercial
provider (GATC, Konstanz, Germany).
Immunofluorescence Assay for Detection of Surface-exposed
BbCRASP-2—For indirect immunofluorescence assays, spirochetes (1
× 10⁷) were incubated with 1:50 dilution of mouse polyclonal
anti-BbCRASP-2 for 1 h at room temperature with gentle agitation. Following
three washes with PBS containing 0.2% BSA, cells were collected by
centrifugation at 14,000 × g for 10 min and resuspended in 100
μl of PBS containing 0.2% BSA. Aliquots of 10 μl were then spotted on
coverslips and allowed to air dry overnight. After fixation with methanol,
samples were dried for 90 min at room temperature and incubated for 90 min in
a humidified chamber with a 1:500 dilution of Alexa 488-conjugated goat
anti-mouse IgG (Molecular Probes). Slides were then washes four times with
0.2% BSA in PBS and mounted in ProLong® Gold Antifade reagent (Molecular
Probes) before being sealed with glass slides. Slides were visualized at a
magnification of × 1,000 using an Olympus CX40 fluorescence microscope
mounted with a DS-5Mc charge-coupled device camera (Nikon).
Immunofluorescence Assay for Detection of Complement
Proteins—Spirochetes (6 × 10⁶) were incubated with
25% NHS for 30 min at 37 °C with gentle agitation, washed three times with
PBS containing 1% BSA (PBS-BSA). Aliquots of 10 μl were then spotted on
microscope slides and allowed to air dry overnight. After fixation, slides
were incubated for 1 h in a humidified chamber with antibodies against
complement components C3 (dilution of 1:1000), C6 (dilution of 1:200), and
C5b-9 (dilution of 1:50). Following four washes with PBS, the slides were
incubated for 1 h at room temperature with 1:2000 dilutions of appropriate
Alexa 488-conjugated secondary antibodies (Molecular Probes, Leiden, The
Netherlands). Slides were then washed, sealed, and visualized as described
above.
Serum Adsorption Experiments—Spirochetes harvested by
centrifugation were resuspended in 500 μl veronal-buffered saline
(supplemented with 1 mm Mg²⁺, 0.15 mm Ca²⁺, 0.1% gelatin, pH 7.4) and a portion of 1 ×
10⁹ organisms were sedimented by centrifugation. The cell sediment
was then resuspended in 750 μl of NHS supplemented with 34 mm EDTA and incubated for 1 h at room temperature with gentle agitation. After
three washes with PBSA (0.15 m NaCl, 0.03 m phosphate,
0.02% sodium azide, pH 7.2) containing 0.05% Tween 20, the proteins bound to
the spirochetes were eluted by incubation with 0.1 m glycine-HCl,
pH 2.0, for 15 min. The bacterial cells were sedimented by centrifugation
(14,000 × g, 20 min, 4 °C), and the proteins in the
supernatant were analyzed by SDS-PAGE and Western blotting.
Serum Susceptibility Testing of Borrelia Strains—Serum
susceptibility of B. garinii strains G1, G1/pKFSS1, G1/pCSPZ, and G1
containing shuttle vector pCSPZ that harbor point mutations in the
cspZ gene was assessed by using a growth inhibition assay as
described previously (7 (link)). Each
experiment was conducted at least three times, and means ± S.D. were
calculated.
Human Sera, CFH, CFHL1, and Monoclonal and Polyclonal
Antibodies—NHS obtained from 20 healthy human blood donors without
known history of spirochetal infections was used as the source for CFH.
Purified human CFH was purchased from Calbiochem. CFHL1 were expressed in
Spodoptera frugiperda Sf9 insect cells infected with recombinant
baculovirus (16 (link),
39 (link)).
Generation of mAb L41 1C11 against FlaB was described elsewhere
(35 ). To detect GST fusion
proteins a goat anti-GST antibody (GE Healthcare, Germany) was used.
Polyclonal rabbit αSCR1–4 antiserum was used for detection of
CFHL1 (16 (link)), and the mAb VIG8
was applied to specifically detect CFH
(35 ). For detection of both
complement regulators a goat anti-human CFH antiserum (Merck Biosciences, Bad
Soden, Germany) was used. The goat anti-human C3 and C6 antibodies were
purchased from Calbiochem, and the monoclonal anti-human C5b-9 antibody was
from Quidel (San Diego, CA). Polyclonal mouse anti-BbCRASP-2 sera were
generated by injection (intraperitoneal) of recombinant BbCRASP-2 into Balb/c
mice (31 (link)).

Siegel C., Schreiber J., Haupt K., Skerka C., Brade V., Simon M.M., Stevenson B., Wallich R., Zipfel P.F, & Kraiczy P. (2008). Deciphering the Ligand-binding Sites in the Borrelia burgdorferi Complement Regulator-acquiring Surface Protein 2 Required for Interactions with the Human Immune Regulators Factor H and Factor H-like Protein 1. The Journal of Biological Chemistry, 283(50), 34855-34863.

Publication 2008

Purification of Bovine Arrestin-3 Truncation

Cited 9 times

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Publication 2011

arrestin3 Bacteria Bos taurus Buffers Cells Cloning Vectors Codon, Terminator DNA, Complementary Gel Chromatography Gold heparin-sepharose Proteins Reading Frames Sepharose Sodium Chloride tris(2-carboxyethyl)phosphine Tromethamine

Purification of RNA Polymerases I and III

Cited 8 times

For purification, 1 kg of cells was suspended in buffer A (250 mM Tris–HCl pH 8, 40% glycerol, 250 mM ammonium sulfate, 1 mM EDTA, 10 mM MgCl₂, 10 µM ZnCl₂, 12 mM β-mercaptoethanol) supplemented with protease-inhibitor cocktail (cOmplete EDTA-free, Roche) and lysed at 4°C with glass beads in a BeadBeater (BioSpec). The soluble fraction obtained after centrifugation (1 h at 14 000 rev min⁻¹ in a Beckmann JA14 rotor) was loaded onto Heparin Sepharose (GE Healthcare) equilibrated in buffer A. The column was washed with buffer B (50 mM Tris–HCl pH 8, 250 mM ammonium sulfate, 0.5 mM EDTA, 1 mM MgCl₂, 10 µM ZnCl₂, 1 mM β-mercaptoethanol, 0.5 mM PMSF) and the complex was eluted from the resin with buffer B* (buffer B with 1 M ammonium sulfate). The sample was diluted to 500 mM ammonium sulfate and incubated with 10 ml pre-equilibrated IgG Sepharose (GE Healthcare) for 6 h. After washing with ten column volumes of buffer C (50 mM Tris–HCl pH 8, 20% glycerol, 225 mM ammonium sulfate, 0.5 mM EDTA, 1 mM MgCl₂, 10 µM ZnCl₂, 2 mM β-mercaptoethanol, 1 mg ml⁻¹ Pefabloc), the IgG beads were mixed with Tobacco etch virus (TEV) protease and incubated overnight at 4°C in the same buffer. The supernatant was recovered and the resin was further washed with ten column volumes of buffer C* (buffer C without glycerol and with only 60 mM ammonium sulfate). The sample was subsequently purified by ion exchange on a Mono Q column (GE Healthcare); elution was performed using a gradient from 60 mM to 1 M ammonium sulfate in buffer D (40 mM Tris–HCl pH 8, 0.5 mM EDTA, 1 mM MgCl₂, 10 µM ZnCl₂, 1 mg ml⁻¹ Pefabloc, 10 mM DTT). Pol I and Pol III eluted at ∼250 and ∼350 mM ammonium sulfate, respectively. The sample was concentrated to 6.5–7 mg ml⁻¹ before crystallization.

Moreno-Morcillo M., Taylor N.M., Gruene T., Legrand P., Rashid U.J., Ruiz F.M., Steuerwald U., Müller C.W, & Fernández-Tornero C. (2014). Solving the RNA polymerase I structural puzzle. Acta Crystallographica Section D: Biological Crystallography, 70(Pt 10), 2570-2582.

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Publication 2014

2-Mercaptoethanol Buffers Cells Centrifugation Crystallization Edetic Acid Glycerin heparin-sepharose immunoglobulin G-sepharose Ion Exchange Magnesium Chloride Mono Q Pefabloc Protease Inhibitors Resins, Plant RNA Polymerase I Sulfate, Ammonium TEV protease Tromethamine

Most recents protocols related to «Heparin-sepharose»

Characterizing TRAIL-Heparin Binding

To characterize the binding of WT TRAIL and TRAIL mutants to heparin, 100 μg of purified WT or mutant TRAIL was applied to a 1 ml HiTrap heparin–Sepharose column (Cytiva Lifesciences) and eluted with a salt gradient from 150 mM to 1 M NaCl at pH 7.1 in 25 mM HEPES buffer. The conductivity measurements at the peak of the elution were converted to the concentration of NaCl based on a standard curve.

Luo Y., Hao H., Wang Z., Ong C.Y., Dutcher R., Xu Y., Liu J., Pedersen L.C, & Xu D. (2024). Heparan sulfate promotes TRAIL-induced tumor cell apoptosis. eLife, 12, RP90192.

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Publication 2024

Characterization of CtsK-Heparin Binding

To characterize the binding of pro-CtsK and mature CtsK to heparin, ~50 μg of purified WT or mutant CtsK was applied to a 1 ml HiTrap heparin–Sepharose column (Cytiva Lifesciences) and eluted with a salt gradient from 150mM to 2M NaCl at pH 5.5 in 20 mM NaOAc buffer. The conductivity measurements at the peak of the elution were converted to the concentration of NaCl based on a standard curve.

Zhang X., Luo Y., Hao H., Krahn J.M., Su G., Dutcher R., Xu Y., Liu J., Pedersen L.C, & Xu D. (2024). Heparan sulfate selectively inhibits the collagenase activity of cathepsin K. bioRxiv.

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Publication Preprint 2024

Affinity Purification of DR5-mTRAIL Complex

Recombinant mouse DR5 (aa53-177)-Fc fusion protein (721-DR, R&D) alone (10 µg), or DR5–mTRAIL complex (10 µg each pre-incubated for 1 hr at room temperature), were loaded onto heparin-Sepharose (Cytiva) gravity column (200 µl bed volume). Column was first washed with 2 ml buffer A (25 mM HEPES, pH7.1, 150 mM NaCl), followed by four elution steps (800 µl each) using buffers containing 300 mM, 500 mM, 1 M, and 2 M NaCl, respectively. 30 µl of eluents from each step were resolved on a 4–20% SDS-PAGE gel and the gel was visualized by silver staining.

Luo Y., Hao H., Wang Z., Ong C.Y., Dutcher R., Xu Y., Liu J., Pedersen L.C, & Xu D. (2024). Heparan sulfate promotes TRAIL-induced tumor cell apoptosis. eLife, 12, RP90192.

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Publication 2024

Heparin Enrichment of Plasma Proteome

The heparin enrichment process was conducted in technical triplicates, using 40 μl of pooled neat human plasma per replicate. Initially, 100 μl of heparin-sepharose bead slurry (1:1 w/v) was prepared with 50 μl of beads for each replicate (n = 3) and binding buffer (50 mM sodium phosphate, pH 7.4). Each slurry was then washed twice with 1 ml of binding buffer. Subsequently, each replicate of neat plasma (40 μl) was diluted with 1 ml of binding buffer to generate diluted plasma (DP) as input and mixed with the heparin-sepharose beads prepared above. The incubation was performed at room temperature for 10 minutes coupled with rotation. Following the enrichment step, the beads were spun down at 500×g for 2 minutes and the supernatant was collected as the heparin-depleted flowthrough (Hp-depleted FT) fraction. The heparin-sepharose beads were then washed twice with 1 ml of binding buffer and resuspended in 1 ml of binding buffer (Hp-enriched fraction). The Hp-enriched fraction was then split into 650 μl for digestion and 350 μl for SDS-PAGE and western blotting. The supernatant was removed before further processing. For Set 1 (n = 36) and Set 2 (n = 85) samples, a global pooled standard (GPS) was prepared for each set as internal control for enrichment by pooling equal amount of each sample within each set. A nearly identical protocol was followed for subsequent enrichment, using 50 μl of beads (Set 1) and 200 μl of beads (Set 2) correspondingly for each set.

Guo Q., Ping L., Dammer E.B., Yin L., Xu K., Shantaraman A., Fox E.J., Golde T.E., Johnson E.C., Roberts B.R., Lah J.J., Levey A.I, & Seyfried N.T. (2024). Heparin-enriched plasma proteome is significantly altered in Alzheimer’s Disease. Research Square.

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Publication Preprint 2024

Preserving CtsK Stability via Heparin Complex

The stability of CtsK assessed by storing either CtsK alone, or CtsK together with heparin (4:1 molar ratio), in a buffer containing 2.5 mM MMTS (a reversible active site inhibitor of cysteine proteases) and 50 mM NaOAc (pH 5.5) at 4 °C for 7 days. On day 1, 2, 3, 5 and 7, ~ 200 ng aliquots were taken to measure the peptidase activity using the fluorogenic substrate as describe above. Also on selected days, 40 µg of CtsK or CtsK/heparin complex were analyzed by heparin-Sepharose chromatography as described above.

Zhang X., Luo Y., Hao H., Krahn J.M., Su G., Dutcher R., Xu Y., Liu J., Pedersen L.C, & Xu D. (2024). Heparan sulfate selectively inhibits the collagenase activity of cathepsin K. bioRxiv.

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Publication Preprint 2024

Top products related to «Heparin-sepharose»

Heparin sepharose by GE Healthcare

Sourced in United States, United Kingdom

Heparin Sepharose is a chromatography resin used for the purification of proteins that interact with heparin, a highly sulfated glycosaminoglycan. It consists of heparin covalently coupled to a cross-linked agarose matrix. The resin can be used to capture and isolate a variety of heparin-binding proteins, including growth factors, enzymes, and coagulation factors.

Heparin sepharose column by GE Healthcare

Sourced in United States

The Heparin-Sepharose column is a chromatography column used for the purification and separation of biomolecules. It contains Sepharose beads covalently linked with the anticoagulant heparin, which serves as a ligand for the affinity-based separation of various proteins, enzymes, and other molecules.

Hitrap heparin hp column by GE Healthcare

Sourced in United States, United Kingdom, Sweden

The HiTrap Heparin HP column is a lab equipment product designed for protein purification. It is a prepacked chromatography column that utilizes heparin, a naturally occurring sulfated glycosaminoglycan, as the ligand for the affinity-based separation of proteins from complex mixtures. The column is intended for use in research and development applications.

Heparin by Merck Group

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Heparin is a pharmaceutical product manufactured by Merck Group. It is a naturally occurring anticoagulant, primarily used as a laboratory reagent to prevent the clotting of blood samples.

Hitrap heparin by GE Healthcare

Sourced in United States, United Kingdom

The HiTrap Heparin is a prepacked chromatography column designed for affinity purification of heparin-binding proteins. It can be used for the purification of various biomolecules, including growth factors, enzymes, and plasma proteins. The column is pre-packed with Sepharose beads coupled with heparin, a sulfated glycosaminoglycan that serves as a ligand for the target proteins.

Hitrap heparin hp by GE Healthcare

Sourced in United States, Sweden

HiTrap Heparin HP is a pre-packed affinity chromatography column designed for the purification of a wide range of heparin-binding proteins, including growth factors, cytokines, and enzymes. The column utilizes immobilized heparin as the ligand, providing a versatile platform for the capture and enrichment of target analytes from complex samples.

Pvdf membrane by Merck Group

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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.

Heparin sepharose 6 fast flow by GE Healthcare

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Heparin Sepharose 6 Fast Flow is a chromatography resin used for the purification of heparin-binding proteins. It consists of heparin coupled to Sepharose 6 Fast Flow beads, providing a high-capacity matrix for the affinity capture of target proteins.

Hitrap heparin sepharose column by GE Healthcare

Sourced in Sweden

The HiTrap heparin-Sepharose column is a pre-packed chromatography column designed for the purification of heparin-binding proteins. It utilizes heparin, a glycosaminoglycan, as the ligand immobilized on Sepharose beads to capture target proteins from complex samples. The column provides a convenient and efficient method for the purification of various heparin-binding proteins, such as growth factors, enzymes, and coagulation factors.

Protease inhibitor cocktail by Merck Group

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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.

More about "Heparin-sepharose"

Heparin-sepharose is a versatile chromatographic material widely used in the purification and analysis of biomolecules, particularly proteins.
It consists of heparin, a sulfated glycosaminoglycan, immobilized on a sepharose matrix.
This technique exploits the affinity of heparin for various proteins, allowing for the selective capture and separation of these molecules based on their interactions with heparin.
Heparin-sepharose chromatography is commonly employed in the study of growth factors, enzymes, and other proteins of biological importance.
It is a powerful tool for researchers investigating protein structure, function, and purification.
Synonyms and related terms include heparin sepharose, heparin-sepharose column, HiTrap heparin HP column, heparin, HiTrap heparin, HiTrap heparin HP, PVDF membranes, heparin sepharose 6 fast flow, HiTrap heparin-sepharose column, and protease inhibitor cocktail.
This versatile technique is widely used in boimolecular research and offers a reproducible and effective approach to protein purification and analysis.