Anti-GFP 抗体 (ab5450)
Key features and details
- Goat polyclonal to GFP
- Suitable for: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-P
- Reacts with: Species independent
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-GFP antibody
GFP 一次抗体 製品一覧 -
製品の詳細
Goat polyclonal to GFP -
由来種
Goat -
アプリケーション
適用あり: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-Pmore details -
種交差性
交差種: Species independent -
免疫原
Recombinant full length protein corresponding to GFP.
Database link: P42212 -
ポジティブ･コントロール
- Pure GFP protein, or cells known to overexpress GFP.
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特記事項
Protein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody.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
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.40
Constituents: 0.79% Tris HCl, 25% Glycerol -
Concentration information loading... -
精製度
Protein G purified -
特記事項（精製）
This antibody is Goat anti-GFP serum (ab5449) affinity purified using a HiTrap-NHS activated sepharose column (Amersham-Pharmacia) containing covalently linked highly purified recombinant GFP. After applying the serum to the column and extensive washing, the eluted anti-GFP immunoglobulins are desalted. -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab5450の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| IP | (1) |
Use at an assay dependent concentration.
|
| ICC |
Use at an assay dependent concentration.
|
|
| IHC-FoFr |
Use at an assay dependent concentration.
|
|
| Electron Microscopy |
Use at an assay dependent concentration.
|
|
| IHC - Wholemount | (2) |
Use at an assay dependent concentration.
|
| ICC/IF | (2) |
1/2000.
|
| WB |
1/2000 - 1/20000.
|
|
| IHC-P | (4) |
1/1000.
|
| 特記事項 |
|---|
|
IP
Use at an assay dependent concentration. |
|
ICC
Use at an assay dependent concentration. |
|
IHC-FoFr
Use at an assay dependent concentration. |
|
Electron Microscopy
Use at an assay dependent concentration. |
|
IHC - Wholemount
Use at an assay dependent concentration. |
|
ICC/IF
1/2000. |
|
WB
1/2000 - 1/20000. |
|
IHC-P
1/1000. |
ターゲット情報
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関連性
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. -
別名
- GFP antibody
- Green fluorescent protein antibody
画像
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image from Sylva M et al., Plos One. 2011;6(10):e26088. Fig 2.; doi: 10.1371/journal.pone.0022616 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemical staining of donor BMDCs expressing GFP in mouse pancreas tissue using ab5450. Antigen retrieval was carried out using target-retrieval solution for 30 minutes in a boiling water bath. Endogenous peroxidase activity was quenched with 3% hydrogen peroxide (5 minutes). Panels A and B were detected with a polymer horseradish peroxidase anti-rabbit detection system (30 minutes) and 3,3′-diaminobenzidine was used as a substrate. Counter-staining was performed with hematoxylin.
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Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)ab5450 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab5450 at 1/2000 dilution overnight at +4°C followed by incubation with ab150129, Donkey Anti-Goat IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab5450 was applied gave a stronger signal in the 488 channel, indicating that ab5450 is binding to GFP and therefore eliciting signal amplification.
ab5450 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image courtesy of an anonymous Abreview.ab5450 staining GFP in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized using 0.2% Triton and then blocked with serum for 1 hour at room temperature, followed by incubation with the primary antibody at a 1/1000 dilution for 14 hours at 4°C. A Cy2®-conjugated secondary antibody was used at a 1/200 dilution. -
IHC - Wholemount - Anti-GFP antibody (ab5450)Image courtesy of Sana Zakaria by Abreview.ab5450 staining GFP in murine hindbrain tissue by IHC-Wholemount.
Dissected hindbrains of E12.5 stage embryos were fixed in 4% paraformaldehyde for 4-5 hours and then washed in PBS before proceeding with immunolabelling in PBS with 1% triton. Primary antibody was incubated for 4-6 days (99-144 hours). The hindbrain was cultured as an explant after GFP was electroporated. The wholemount immuno with ab5450 at a 1/500 dilution was done to see if GFP could be detected, it was the same GFP expression with the antibody as that which had been seen initially with the electroporation. The secondary used was an Alexa-Fluor 647 conjugated donkey anti-goat polyclonal used at a 1/150 dilution. Blue is DAPI and green is the GFP antibody (pseudo-colored from red Alexa Fluor 647). -
Electron Microscopy - Anti-GFP antibody (ab5450)This image is courtesy of an Abreview submitted by Narciso CamposElectron Microscopy of Arabidopsis thaliana tissue sections labelling GFP with ab5450. An 18nm gold-conjugated Donkey anti-goat IgG polyclonal (1/15) was used as the secondary antibody.
Arabidopsis thaliana transgenic plant expressing GFP fused to an Endoplasmic Recticulum (ER) marker. No label is observed in: C - chloroplasts, M - mitochondria, G - golgi. The sample was prepared by cryofixation and embedding in Lowicryl HM20 resin. The image was taken in a JEOL 1010 transmission electron microscope at 20,000× magnification.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (172)
ab5450 は 172 報の論文で使用されています。
- Vastagh C et al. Networking of glucagon-like peptide-1 axons with GnRH neurons in the basal forebrain of male mice revealed by 3DISCO-based immunocytochemistry and optogenetics. Brain Struct Funct 226:105-120 (2021). PubMed: 33169188
- Kahn BM et al. The vascular landscape of human cancer. J Clin Invest 131:N/A (2021). PubMed: 33258803
- Lafferty RA et al. Positive Effects of NPY1 Receptor Activation on Islet Structure Are Driven by Pancreatic Alpha- and Beta-Cell Transdifferentiation in Diabetic Mice. Front Endocrinol (Lausanne) 12:633625 (2021). PubMed: 33716983
- Pavel M et al. a-Catenin levels determine direction of YAP/TAZ response to autophagy perturbation. Nat Commun 12:1703 (2021). PubMed: 33731717
- Cleal L et al. Resolving the heterogeneity of diaphragmatic mesenchyme: a novel mouse model of congenital diaphragmatic hernia. Dis Model Mech 14:N/A (2021). PubMed: 33735101