Anti-GFP抗体(ab13970)
Key features and details
- Chicken polyclonal to GFP
- Suitable for: WB, ICC/IF
- Reacts with: Species independent
- Isotype: IgY
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术,可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
概述
-
产品名称
Anti-GFP抗体
参阅全部 GFP 一抗 -
描述
鸡多克隆抗体to GFP -
宿主
Chicken -
特异性
Our GFP antibody does cross-react with the many fluorescent proteins that are derived from the jellyfish Aequorea victoria. These are all proteins that differ from the original GFP by just a few point mutations (EGFP, YFP, mVenus, CFP, BFP etc.).
-
经测试应用
适用于: WB, ICC/IFmore details -
种属反应性
与反应: Species independent -
免疫原
Recombinant full length protein corresponding to GFP.
Database link: P42212 -
阳性对照
- ICC: GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line). WB: Transgenic mouse spinal cords.
-
常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate -
Concentration information loading...
-
纯度
IgY fraction -
纯化说明
Sterile filtered. -
克隆
多克隆 -
同种型
IgY -
研究领域
相关产品
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab13970于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
WB | (13) |
1/5000.
|
ICC/IF | (22) |
1/2000.
Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
说明 |
---|
WB
1/5000. |
ICC/IF
1/2000. Used at a dilution of 1/2000 for 1 hr (see Abreview for further information). |
靶标
-
相关性
Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.
Subunit structure: Monomer.
Tissue specificity: Photocytes.
Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.
Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.
Sequence similarities: Belongs to the GFP family.
Biophysicochemical properties: Absorption: Abs(max)=395 nm
Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm. -
别名
- GFP antibody
- Green fluorescent protein antibody
图片
-
All lanes : Anti-GFP antibody (ab13970) at 1/2000 dilution (Diluent 1x TBS /4 hours at 4°C)
Lane 1 : 3 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lane 2 : 2 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lane 3 : 1 µg of GFP plasmid overexpressed in mouse cardiomyocytes whole cell lysate with BSA / for 1 hour at room temperature
Lysates/proteins at 25 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (ab150176) at 1/5000 dilution
Performed under reducing conditions.
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 secondsGel Running Conditions: Reduced Denaturing (15% PAGE)
Detection method: Fluorescent Secondary Antibodies
-
ab13970 staining GFP in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF.
Cells were paraformaldehyde fixed, permeabilized with 0.5% triton and blocked with 2% antibody dilution buffer for 2 hours. Cells were incubated with the primary antibody (1/1000) for 1 hour at 25°C. An undiluted Alexa Fluor® 488 conjugated Goat anti-chicken polyclonal was used as the secondary antibody.
-
All lanes : Anti-GFP antibody (ab13970) at 1/1000 dilution
Lane 1 : Whole cell lysate prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
Lane 2 : Whole cell lysate prepared from HeLa cells.
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : IRDye 800CW conjugated goat anti-chicken polyclonal at 1/15000 dilution
-
Transgenic mice expressing GFP selectively in lamina II of the spinal cord.
In the right panels, note the correspondance between the green (rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam) indicating that these two antibody preparations recognized the same gene product. The secondary antibody used with ab13970 was an FITC-labeled goat anti-chicken
-
ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line) cells.
The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab13970 at 1/2000 dilution overnight at +4°C followed by incubation with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1 μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH/3T3 cells, the cells upon which ab13970 was applied gave a stronger signal in the 488 channel, indicating that ab13970 is binding to GFP and therefore eliciting signal amplification.
ab13970 was also applied to non-GFP-transfected NIH/3T3 cells, which produced no positive staining, indicating specificity for GFP.Nuclear DNA was labeled with 1.43 μM DAPI (blue).
-
Lane 1 : Rabbit anti-GFP
Lane 2 : Anti-GFP antibody (ab13970)
All lanes : Transgenic mouse spinal cordsWestern blot of transgenic mouse spinal cords showing that the rabbit anti-GFP (lane 1) and the chicken anti-GFP (Abcam; lane 2) recognize a band at the same molecular weight.
-
ab13970 staining GFP + tumor in mouse muscle cells by ICC/IF.
Cells were formaldehyde fixed and blocked with 3% BSA for 1 hour at 24°C prior to incubation with the primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.
-
Immunocytochemical/ Immunoflurescence analysis of cytospined HEK-293 cells (Human epithelial cell line from embryonic kidney) transfected with GFP, labeling GFP with ab13970 at 1/200 incubated for 16 hours at 4°C with 1% BSA in PBS. Secondary used was a donkey anti-chicken polyclonal DyLight® 594 at 1/500.
GFP is shown in red (DyLight® 594).
Nuclei are counterstained in blue (DAPI).
The left panel shows HEK-293 cells transfected with GFP and the right panel shows non-transfected HEK-293 cells.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (3610)
ab13970 被引用在 3610 文献中.
- Szewczyk LM et al. Astrocytic β-catenin signaling via TCF7L2 regulates synapse development and social behavior. Mol Psychiatry 29:57-73 (2024). PubMed: 37798419
- Pallucchi I et al. Molecular blueprints for spinal circuit modules controlling locomotor speed in zebrafish. Nat Neurosci 27:78-89 (2024). PubMed: 37919423
- Hofstetter J et al. Spt5 interacts genetically with Myc and is limiting for brain tumor growth in Drosophila. Life Sci Alliance 7:N/A (2024). PubMed: 37935464
- Zhuo Y et al. Improved green and red GRAB sensors for monitoring dopaminergic activity in vivo. Nat Methods 21:680-691 (2024). PubMed: 38036855
- Eley L et al. eNOS plays essential roles in the developing heart and aorta linked to disruption of Notch signalling. Dis Model Mech 17:N/A (2024). PubMed: 38111957