Anti-GFP抗体
参阅全部 GFP 一抗
山羊多克隆抗体to GFP
Goat
适用于: IP, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-Pmore details
与反应: Species independent
Recombinant full length protein corresponding to GFP.
Database link: P42212
Pure GFP protein, or cells known to overexpress GFP.
Protein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody.
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Liquid
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.
pH: 7.40
Constituents: 0.79% Tris HCl, 25% Glycerol
浓度
25 µg 浓度为 0.5 mg/ml
Protein G purified
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.
多克隆
IgG
Abpromise™承诺保证使用ab5450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 |
|---|---|---|
| IP | (1) | 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. |
GFP antibody
Green fluorescent protein antibody
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.
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 Campos
Electron 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.
