VECTOR SBA生物素标记的大豆凝集素SBA Detection of Glycoproteins using Lectins in Histochemistry,ELISA, and Western Blot Applications
详细介绍
VECTOR SBA生物素标记的大豆凝集素
isolated from Glycine max （soybean） seeds
Composed of four subunits of approximay equal size, soybean agglutinin is a family of closely related isolectins. This glycoprotein has a molecular weight of about 120,000 and an isoelectric point near pH 6.0. SBA preferentially binds to oligosaccharide structures with terminal a- or b-linked N-acetylgalactosamine, and to a lesser extent, galactose residues. Binding can be blocked by substitutions on penultimate sugars, such as fucose attached to the penultimate galactose in blood group B substance. SBA has been used in glycoprotein fractionation, histochemical applications and cell sorter analysis.
An important application for SBA is the separation of pluripotential stem cells from human bone marrow. Cells fractionated by SBA do not produce graft vs host disease and can be used in bone marrow transplantation across histocompatibility barriers （references available upon request）. It should be noted that some forms of SBA seem to be excellent in separating human cells while others are better for cells of other species.
VECTOR SBA生物素标记的大豆凝集素
This biotinylated lectin conjugate is prepared from affinity-purified lectin and is optimally labeled with biotin. Essentially free of inactive lectin conjugate and containing no free biotin, this biotinylated lectin provides an ideal intermediate for examining glycoconjugates using the Biotin-Avidin System. First the biotin-labeled lectin is added, followed by the VECTASTAIN® ABC Reagent, Avidin D conjugate, or streptavidin derivative. Another possible application is in the isolation of lymphokines and other products of mitogenic stimulation.
Inhibiting/Eluting Sugar: 200 mM N-acetylgalactosamine
GEwhatman597号折叠滤纸10311845GEwhatman定性折叠滤纸597号折叠滤纸10311845定性滤纸――预折叠级 Whatman定性滤纸为使用提供了方便，节省了时间，相对 普通平整滤纸，它的优点是： l 对于重复性或者多重分析，预折叠滤纸节省了折叠成适合 形状的时间。 l 由于更多表面的暴露，可以减少整个过滤时间。一般，过 滤速度的下降是由于颗粒堆积而产生。 l 更多过滤面积使得整体载量也有所上升。 l 折叠后，自我支撑能力的上升，减少同漏斗接触有利于维 持流速。 l 预折叠并不会显著影响技术参数，数值同相应的平整滤纸 一样。
核酸研究 碱洗提和DNA段分馏法 海洋学研究 透明的聚碳酸酯膜是浮游生物研究的一个新工具。这些超薄的透明滤膜虽然柔韧但强度非常好，可以过江浮游生物样品并直接安装奔玻片上进行观察。（Ref:Hewes et al.1998; Graham and Mitchell 1999; Graham 1999.） 医疗器械和体外诊断 生物传感器-作为生物试剂和电化学探针的控制隔离 诊断分析-控制流速、样品制备、血液分离和乳胶微粒的捕捉 细胞生物学-细胞培养、趋化现象和细胞学分的，例如直接着色、同位素和荧光分析。 皮肤吸收药物的载体-治疗药物的惰性载体。
Biotinylated Pisum Sativum Agglutinin豌豆凝集素Detection of Glycoproteins using Lectins in Histochemistry,ELISA, and Western Blot Applications
详细介绍
Biotinylated Pisum Sativum Agglutinin豌豆凝集素
isolated from Pisum sativum （garden pea） seeds Pisum sativum agglutinin is nearly identical in structure and carbohydrate specificity to Lens culinaris agglutinin. PSA has four subunits, two of approximay 17,000 daltons and two of about 6,000 daltons. Isoelectric focusing has revealed two isolectins with isoelectric points of pH 5.9 and pH 7.0. The lectin has specificity toward a-linked mannose-containing oligosaccharides, with an N-acetylchitobiose-linked a-fucose residue included in the receptor sequence. Calcium and manganese ions are required for activity. PSA has been used to fractionate cells, to isolate glycoproteins and glycopeptides, to distinguish between normal and virally transformed cells, as a T-cell mitogen, and as an inhibitor of allograft rejection.
Biotinylated Pisum Sativum Agglutinin豌豆凝集素
This biotinylated lectin conjugate is prepared from affinity-purified lectin and is optimally labeled with biotin. Essentially free of inactive lectin conjugate and containing no free biotin, this biotinylated lectin provides an ideal intermediate for examining glycoconjugates using the Biotin-Avidin System. First the biotin-labeled lectin is added, followed by the VECTASTAIN® ABC Reagent, Avidin D conjugate, or streptavidin derivative. Another possible application is in the isolation of lymphokines and other products of mitogenic stimulation. Inhibiting/Eluting Sugar: 200 mM a-methyl mannoside/200 mM a-methyl glucoside mixture
怀槐凝集素Biotinylated Maackia Amurensis LectinBiotinylated Maackia Amurensis Lectin (MAL) II Detection of Glycoproteins using Lectins in Histochemistry,ELISA, and Western Blot Applications
详细介绍
怀槐凝集素Biotinylated Maackia Amurensis Lectin
Detection of Glycoproteins using Lectins in Histochemistry, ELISA, and Western Blot Applications The following protocols offer guidelines for assay development using lectin-based detection of glycoproteins present in tissue sections, adsorbed onto microtiter plates, or transferred from electrophoretic gels onto nitrocellulose or PVDF membranes. Histochemistry: 1a. Staining procedure for paraffin sections: Deparaffinize and hydrate tissue sections through xylenes or other clearing agents and graded alcohol series and rinse for 5 minutes in tap water. If required, retrieve antigens using the Antigen Unmasking Solution （H-3300 or H-3301）. 1b. Staining procedure for frozen sections: Air dry sections. Immediay before staining, fix sections with acetone. Transfer slices to buffer. If endogenous enzyme activities are present, inactivate using appropriate methods. 2. Perform Streptavidin/Biotin blocking if required following kit instructions （SP-2002）. Do not use SP-2001. Block non-specific binding by incubating section with Carbo-Free™ Blocking Solution （Cat. No. SP-5040） for 30 minutes at room temperature. Blot excess blocking solution from the sections. 3. Apply biotinylated lectin at approximay 2-20 μg/ml in PBS （10 mM sodium phosphate, 150 mM NaCl, pH 7.4） to the sections and incubate for 30 minutes at room temperature. Wash with TPBS （PBS + 0.05% Tween™20）. 4. Prepare VECTASTAIN® ® ABC （peroxidase, Cat. No. PK-6100） or VECTASTAIN® ABC-AP （alkaline phosphatase, Cat. No. AK-5000） reagents according to the kit instructions. Apply to the sections and incubate for 30 minutes at room temperature. Wash with TPBS. 5. Apply an appropriate precipitating substrate for the enzyme system used in step 4. For peroxidase, ImmPACT™ DAB （Cat. No. SK-4105） is recommended; for alkaline phosphatase, Vector® Red （Cat. No. SK-5100）. Rinse in tap water. 6. Counterstain （optional）, clear and mount. For galactose or GalNAc-specific lectins avoid mounting in glycerol-based mounting media. ELISA: 1. Adsorb target protein to microtiter plate by placing 50-200 μl of approximay 3 μg/ml glycoprotein solution into the desired wells. Some wells may be left untreated as negative controls. Incubate at 37 ºC for 1 hour. Wash wells three times with TPBS （PBS + 0.05% Tween™20）. 2. Block non-specific binding by filling each well to the brim with Carbo-Free™ Blocking Solution （Cat. No. SP-5040） for 30 minutes at room temperature. Wash wells three times with TPBS. 3. Apply 50-200 μl of approximay 2-20 μg/ml biotinylated lectin in PBS to the wells and incubate for 30 minutes at room temperature. Wash wells three times with TPBS. Vector Laboratories, Inc., 30 Ingold Road, Burlingame, CA 94010 U.S.A.
怀槐凝集素Biotinylated Maackia Amurensis Lectin
VECTOR LABORATORIES VECTOR LABORATORIES 4. Prepare VECTASTAIN® ® ABC （peroxidase, Cat. No. PK-6100） or VECTASTAIN® ABC-AP （alkaline phosphatase, Cat. No. AK-5000） reagents according to the kit instructions. Apply to the wells and incubate for 30 minutes at room temperature. Wash wells three times with TPBS. 5. Apply an appropriate non-precipitating substrate for the enzyme system used in step 4. For peroxidase, ABTS （Cat. No. SK-4500） is recommended; for alkaline phophatase, pNPP （Cat. No. SK-5900）. 6. Quantify the colored reaction product by spectrophotometry. Western Blot: 1. Perform electrophoresis and transfer proteins to a membrane according to standard procedures. 2. Block non-specific binding by incubating the membrane in Carbo-Free™ Blocking Solution （Cat. No. SP-5040） for 30 minutes at room temperature. Use a sufficient volume to compley cover the membrane. 3. Incubate membrane in PBS containing approximay 2-20 μg/ml biotinylated lectin for 30 minutes at room temperature. Wash with TPBS （PBS +0.05% Tween™20）. 4. Prepare VECTASTAIN® ® ABC （peroxidase, Cat. No. PK-6100） or VECTASTAIN® ABC-AP （alkaline phosphatase, Cat. No. AK-5000） reagents according to the kit instructions. Incubate the membrane in the reagent for 30 minutes at room temperature. Wash with TPBS. 5. Apply an appropriate substrate for the enzyme system used in step 4. For peroxidase, DuoLuX™ Chemiluminescent/Fluorescent Substrate for Peroxidase （Cat. No. SK-6604） or ImmPACT™ DAB （Cat. No. SK-4105） are recommended; for alkaline phosphatase, Chemiluminescent/Fluorescent Substrate for Alkaline Phosphatase （Cat. No. SK-6605） or BCIP/NBT （Cat. No. SK-5400） are recommended. Negative Controls Negative controls should be run in parallel in each of the above described methodologies to validate binding results. When applying lectins, one of the most appropriate negative controls is to preabsorb the lectin with a concentration of a defined sugar, with which, the lectin has a known high affinity. Vector Labs offers a series of sugars that are intended for such a purpose. The lectin is diluted to a suitable working concentration in a solution containing approximay 200 mM to 500 mM of the sugar. This mixture is left to bind at room temperature for 30 to 60 min. Following this absorption incubation, the mixture is substituted into the procedure in place of the unabsorbed lectin and incubated under the same conditions. The subsequent detection procedure is followed as for the test method. In most cases the vast majority of lectin binding to the tissue section （membrane blot, etc.） will be eliminated. Some trace binding to the section （blot etc） may still be present under these conditions and probably indicates presence of secondary or tertiary sugar preferences. These negative control results should be compared with the test results to determine specificity of binding
This is a 90mm Hydrophobic PTFE membrane filter with a 5.0µm pore size. Mitex is an unsupported hydrophobic PTFE membrane. It can be used under extreme chemical or temperature conditions that other membranes cannot withstand (up to 260 °C).
