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Rabbit抗原名称
GFP分子别名
Green fluorescent proteinAccession
P42212克隆号
S-296-32抗体类型
Rabbit mAb应用
ICFCM, ChIP, ICC, WB, IP交叉反应
This antibody is reactive against all variants of Aequorea victoria GFP such as CFP, ECFP, YFP, BFP, EEGFP, Clover, EYFP and EVenus.
纯化方式
Protein A浓度
0.5 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
储存条件
12 months from date of receipt / reconstitution, -20 °C as supplied
| 应用 | 稀释度 |
|---|---|
| WB | 1:1000 |
| IP | 1:50 |
| ICC | 1:500 |
| ICFCM | 1:5000 |
| ChIP | 1:20-1:50 |
The green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range.[PMID: 28749] The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP.
免疫印迹
WB result of GFP Rabbit mAb
Primary antibody: GFP Rabbit mAb at 1/5000 dilution
Lane 1: 293T whole cell lysate 5 µg
Lane 2: 293T transfected with GFP-tag-fusion protein whole cell lysate 5 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 28 kDa
Observed MW: 28 kDaWB result of GFP Rabbit mAb
Lysate: Aequorea victoria GFP Protein 20ng
Primary antibody:
Lane 1: GFP Rabbit mAb at 1/5000 dilution
Lane 2: GFP Rabbit mAb at 1/10000 dilution
Lane 3: GFP Rabbit mAb at 1/50000 dilution
Lane 4: GFP Rabbit mAb at 1/100000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 28 kDa
Observed MW: 28 kDa
流式分析
Flow cytometric analysis of 4% PAF fixed 90% methanol permeabilized GFP-transfected 293T cells labelling GFP antibody at 1/5000 (0.01 μg) dilution / (right panel) compared with a Rabbit IgG Isotype Control / (left panel). Goat Anti-Rabbit IgG Alexa Fluor 647 was used as the secondary antibody.
免疫沉淀
GFP Rabbit mAb at 1/50 dilution (0.5 µg) immunoprecipitating GFP in 0.2 mg 293T transfected with GFP whole cell lysate.
Western blot was performed on the immunoprecipitate using GFP Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1: 293T transfected with GFP whole cell lysate 5 µg (Input)
Lane 2: GFP Rabbit mAb IP in 293T transfected with GFP whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in 293T transfected with GFP whole cell lysate
Predicted MW: 27 kDa
Observed MW: 27 kDa
免疫细胞化学
ICC shows positive staining in GFP-transfected HeLa cells. Anti-GFP antibody was used at 1/500 dilution (Red) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
Negative control: ICC shows negative staining in Vector-transfected HeLa cells. Anti-GFP antibody was used at 1/500 dilution and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
ChIP
Chromatin immunoprecipitation (ChIP) was
performed on 293T cells were either untransfected
(left panel) or transfected with an GFP-tagged human
H3 construct (right panel) cross - linked with 1%
formaldehyde for 10 min, then chromatin was
fragmented by sonication.
Parallel reactions used GFP Recombinant Rabbit mAb
(S-296-169), Histone H3 Recombinant Rabbit mAb
(SDT-266-44) and Rabbit mAb IgG Isotype
Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed,
eluted, and cross - links reversed. Purified DNA was
analyzed by qPCR.
qPCR (%input: immunoprecipitated DNA/input DNA)
showed the enrichment of RPL30, MYOD1 and SAT-2 in
GFP Recombinant Rabbit mAb (S-296-169)-
immunoprecipitated sample.
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