Anti-GFP抗体[9F9.F9] (ab1218)
![Western blot - Anti-GFP antibody [9F9.F9] (ab1218)](https://www.abcam.cn/ps/products/1/ab1218/Images/ab1218-263056-1218wb.JPG "Western blot - Anti-GFP antibody [9F9.F9] (ab1218)")
Key features and details
- Mouse monoclonal [9F9.F9] to GFP
- Suitable for: WB, IHC-Fr, Sandwich ELISA, ICC/IF
- Reacts with: Species independent
- Isotype: IgG1
概述
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产品名称
Anti-GFP抗体[9F9.F9]
参阅全部 GFP 一抗 -
描述
小鼠单克隆抗体[9F9.F9] to GFP -
宿主
Mouse -
特异性
Assayed by ELISA for direct binding of antigen recognizes wild type and recombinant GFP. Well suited to titrate GFP in solution using either form of the antibody as the capture or detection antibodies. Shows no reactivity against red fluorescence protein (RFP). This antibody is known to cross react with the wild type (wt), recombinant (rGFP) and enhanced (eGFP) forms. -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF Species independentWB Species independent
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免疫原
Fusion protein corresponding to GFP aa 1-246. Full Length Fusion Protein. Derived from the jellyfish Aequorea victoria.
Database link: P42212 -
阳性对照
- Pure GFP protein, or cells known to overexpress GFP.
性能
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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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate -
Concentration information loading... -
纯度
Protein A purified -
纯化说明
GFP Monoclonal Antibody was prepared from tissue culture supernatant by Protein A affinity chromatography. -
克隆
单克隆 -
克隆编号
9F9.F9 -
同种型
IgG1 -
轻链类型
kappa -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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sELISA pair antibody
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab1218 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| ICC/IF |
Species independent
|
| WB |
Species independent
|
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (21) |
1/3000 - 1/30000.
(See Abreviews). Recognises the immunogen under native conditions and reduced/denatured conditions. |
| IHC-Fr | (2) |
1/1000 - 1/5000.
Works also with eYFP. |
| Sandwich ELISA |
1/80000 - 1/500000. Can be paired for Sandwich ELISA with Biotin Goat polyclonal to GFP (ab6658).
ab1218 can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP is well suited to titrate GFP in a sandwich ELISA in combination with polyclonal anti-GFP as a capture antibody. |
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| ICC/IF | (12) |
Use at an assay dependent concentration.
|
| 说明 |
|---|
|
WB
1/3000 - 1/30000. (See Abreviews). Recognises the immunogen under native conditions and reduced/denatured conditions. |
|
IHC-Fr
1/1000 - 1/5000. Works also with eYFP. |
|
Sandwich ELISA
1/80000 - 1/500000. Can be paired for Sandwich ELISA with Biotin Goat polyclonal to GFP (ab6658). ab1218 can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP is well suited to titrate GFP in a sandwich ELISA in combination with polyclonal anti-GFP as a capture antibody. |
|
ICC/IF
Use at an assay dependent concentration. |
靶标
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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
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Western blot - Anti-GFP antibody [9F9.F9] (ab1218)All lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1 mg/ml (1 hour at room temperature)
Lane 1 : HeLa lysate 50ng with BLOTTO, overnight at 4°C
Lane 2 : HeLa lysate 100ng with BLOTTO, overnight at 4°C
Lane 3 : HeLa lysate 500ng with BLOTTO, overnight at 4°C
Blocking peptides at 5 % per lane.
Secondary
All lanes : IRDye® 800 conjugated Goat anti-mouse IgG (H+L), 45 minutes at room temperature at 1/2500 dilution
Additional bands at: 27 kDa. We are unsure as to the identity of these extra bands. -
![Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [9F9.F9] (ab1218)](https://www.abcam.cn/ps/products/1/ab1218/Images/ab1218-350041-anti-gfp-antibody-9f9f9-immunocytochemistry-immunofluorescence-human-hela.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [9F9.F9] (ab1218)This image is courtesy of an anonymous AbreviewImmunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling GFP with ab1218 at 1/100. Cells were fixed with Formaldehyde and permeabilised with 0.2% Triton100 (permeablized after primary incubation). A good anti-mouse Texas Red® at 1/1000 was used as the secondary antibody. -
![Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [9F9.F9] (ab1218)](https://www.abcam.cn/ps/products/1/ab1218/Images/ab1218-318508-1218.JPG)
Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [9F9.F9] (ab1218)Grant, M.P. et al PLoS One. 2015 Aug 28;10(8):e0136702. doi: 10.1371/journal.pone.0136702. eCollection 2015TIRFM responses to prolonged stimulation with 10 mM Ca2+
C. Cells expressing BS-wt were stimulated with 0.5 or 10 mM Ca2+ for 60 min, 37°C, followed by fixation, permeabilization, and staining with anti-GFP antibody (secondary antibody tagged with Alexa-568). Confocal images of SEP and anti-GFP fluorescence are shown along with the merged image. D. Experiment as in C for cells expressing BS-5A mutant.
HEK-293 cells were seeded on glass coverslips and transiently transfected with BS-wt or BS-5A cDNAs. Cells were incubated in DMEM containing 0.5 or 10 mM Ca2+ for 1 hr at 37°C, 5% CO2. Following incubation, cells were immediately fixed in 4% paraformaldehyde for 10 min, RT, permeabilized with 0.2% Triton X-100 (10 min), blocked with 5% serum and 0.1% BSA then incubated with monoclonal anti-GFP antibody (Abcam), followed by goat anti-mouse secondary antibody conjugated to Alexa-568.
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![Western blot - Anti-GFP antibody [9F9.F9] (ab1218)](https://www.abcam.cn/ps/products/1/ab1218/Images/ab1218-263054-ab1218wb.jpg)
Western blot - Anti-GFP antibody [9F9.F9] (ab1218)Image courtesy of an anonymous AbreviewAll lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1/1000 dilution (2 hours at 25°C)
Lane 1 : J774 with Milk, 2 hours at 25°C
Lane 2 : J774+pEGFP-N1 with Milk, 2 hours at 25°C
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat anti-mouse HRP at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Additional bands at: 26 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
实验方案
文献 (289)
ab1218 被引用在 289 文献中.
- Zhang Q et al. Multisite Phosphorylation Determines the Formation of Ska-Ndc80 Macro-complexes That Are Essential for Chromosome Segregation during Mitosis. Mol Biol Cell N/A:mbcE19100569 (2020). PubMed: 32491969
- Binder JL et al. Optical induction of autophagy via Transcription factor EB (TFEB) reduces pathological tau in neurons. PLoS One 15:e0230026 (2020). PubMed: 32208437
- Finke J et al. Anatomy of a viral entry platform differentially functionalized by integrins a3 and a6. Sci Rep 10:5356 (2020). PubMed: 32210347
- Patel AA et al. Simultaneous Electrophysiology and Fiber Photometry in Freely Behaving Mice. Front Neurosci 14:148 (2020). PubMed: 32153363
- Mason-Osann E et al. RAD54 promotes alternative lengthening of telomeres by mediating branch migration. EMBO Rep 21:e49495 (2020). PubMed: 32337843
