欧美三级久久三级久久三级,喷喷喷黄色视频,嗯~阿插进去深一点无码,亚洲欧美一区二区三区在线

您好,歡迎進(jìn)入上海力敏實(shí)業(yè)有限公司網(wǎng)站!
一鍵分享網(wǎng)站到:
您現(xiàn)在的位置:首頁(yè) >> 產(chǎn)品中心 >> 生化試劑、抗體、血清 >> 常規(guī)生化試劑 >> B-1245VECTOR GNL生物素雪花蓮凝集素Biotinylated Galanthus Nivalis

VECTOR GNL生物素雪花蓮凝集素Biotinylated Galanthus Nivalis

  • 更新時(shí)間:  2023-07-24
  • 產(chǎn)品型號(hào):  B-1245
  • 簡(jiǎn)單描述
  • VECTOR GNL生物素雪花蓮凝集素Biotinylated Galanthus Nivalis Lectin
    Detection of Glycoproteins using Lectins in Histochemistry,
    ELISA, and Western Blot Applications
詳細(xì)介紹

VECTOR GNL生物素雪花蓮凝集素Biotinylated Galanthus Nivalis Lectin

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.

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

 

上海力敏實(shí)業(yè)有限公司代理VECTOR產(chǎn)品,咨詢(xún)VECTOR GNL生物素雪花蓮凝集素Biotinylated Galanthus Nivalis Lectin 詳細(xì)產(chǎn)品信息


留言框

  • 產(chǎn)品:

  • 您的單位:

  • 您的姓名:

  • 聯(lián)系電話:

  • 常用郵箱:

  • 省份:

  • 詳細(xì)地址:

  • 補(bǔ)充說(shuō)明:

  • 驗(yàn)證碼:

    請(qǐng)輸入計(jì)算結(jié)果(填寫(xiě)阿拉伯?dāng)?shù)字),如:三加四=7
骚妇吃鸡巴视频| 天天视频一二三四区入口| 久久精品国产亚洲av天海| 农民工猛吸女大学奶头| 黄色视频啊嗯嗯快点进来| 女人张开腿让男人桶视频| 日韩一区二区肥| 大陆日逼小视频| 欧亚乱熟女一区二区在线| 国产精品色婷婷在线观看| 男人插女人喷水视频网站| 男人j插女人j黄色网站| 大阴经网站熟女毛片视频| 精品无码国产污污污免费网站2| 成人毛片女人毛片免费看| 色噜噜久久一区二区三区| 久久久999久久久久久| 九九热视频精品在线观看| 欧美潮喷在线| 肏胖老太太视频| 东北妇女操逼网| 东北女人大黑逼| 男人和女人肏屄黄色网站| 就去吻亚洲精品日韩都没| 欧美性激烈粗大精品xxx| 女人曰大逼视频| 国产真实熟女内射精品视频| 国产精品毛片无码无遮拦| 国产一级特黄a大片99| 日本亚洲成人中文激情网| 任我爽精品视频在线观看| 日韩中文字幕丝袜办公室| 午夜国产羞羞羞国产精品| 国产精品久久91xxx| 国产高清熟女在线视频播放| 大香蕉在线网大香蕉在线| 极品尤物一区二区三区| 色妞av一区二区在线观看| 在线免费观看日本中文字幕| 九一麻豆蜜臀一区二区三区| 久久99精品久久久久婷婷|