Antigen Information
- P23769
- 2624
- GATA2
- Human
Assay Format
Product Specifications
Introduction
Product Features
- Specific transcription factor-DNA binding assay
- Perfect alternative to EMSA
- Easy to perform in an ELISA format
- Non-radioactive assay
- High throughput (96-well plate format)
- Assay can be completed within 5 hours
Application Notes
- 96-well Strip Microplate pre-coated with DNA probes
- DNA Binding Buffer
- Positive Control Sample
- Specific Competitor DNA probe
- Non-specific Competitor DNA probe
- Assay Reagent
- DTT
- Wash Buffer
- Primary Antibody
- HRP-conjugated Secondary Antibody
- Antibody Diluent Buffer
- TMB One-Step Substrate Reagent
- Stop Solution
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Absorbent paper
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation < li="">
- Microplate reader capable of measuring absorbance at 450 nm
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared HRP-secondary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Typical Data
Figure 1Transcription factor activity assay of GATA-2 from nuclear extracts of K562 cells or HeLa cells. A. Western-blot result of GATA-2 from cytoplasmic and nuclear fractions. B. Transcription factor activity assay of GATA-2 from nuclear fractions with the RayBio® GATA-2 TF-Activity Assay Kit.
Figure 2Transcription factor activity assay of GATA-2 from nuclear extracts of K562 cells or HeLa cells with the specific competitor or non-specific competitor. The result shows specific binding of GATA-2 to the GATA conserved binding site detected by using the RayBio® GATA-2 TF-Activity Assay Kit.
Storage/Stability
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1.小鼠Lewis肺癌细胞DNA含量测定方法
(1).从C57BL/6小鼠上切除肿块,在培养皿内用PBS冲洗.
(2).去除结缔组织及脂肪,剪碎肿块.
(3).小碎片移入1.20×38mm注射针,加压使其通过,于4℃条件下重悬细胞于HBSS中.
(4).将200~300μL细胞悬液(5×105细胞/mL)中加入3mL PI(50μg/mL),染色3LL细胞,于4℃存放20~30分钟.
(5).测定580~750nm之间的发射荧光,以去除末结合PI产生的激发光与发射光谱线之间的重叠部分.
请各位大侠给予帮助!!
谢谢!!
希望同路人多多交流啊!