Green Fluorescent Protein antibody
Sheep anti Green Fluorescent Protein antibody used for the identification of GFP tagged VEGF-A expressing cells by immunofluorescence.
Image caption:
HSV-1 infection drives transcriptional upregulation of VEGF-A.
(A) Representative image of a cornea from pVEGFA-GFP reporter mice at day 3 PI with HSV-1 strain McKrae stained for HSV-1 antigen (red), GFP transcriptional reporter for VEGF-A (green) and DAPI (blue) (B) GFP transcriptional reporter expression (green) with HSV-1 antigen (red) at 12 hours PI with HSV-1 strain McKrae. (C) Real time PCR of THCE cells at 12 hours PI with 3 pfu per cell HSV-1 strain McKrae with VEGF-A fold induction determined via the geometric means of VEGF-A induction relative to the housekeeping genes β-actin, TBP and PPIA (** p<0.01). (D) Real time PCR of VEGF-A transcription in 293 cells infected with 3 pfu per cell HSV-1 strain McKrae also determined via geometric mean relative to β-actin, TBP and PPIA (** p<0.01) and (E) Levels of VEGF-A by cytokine bead array in cytoplasmic extracts of 293 cells at the indicated times PI with 3 pfu per cell HSV-1 strain McKrae, expressed as pg of VEGF-A per μg of cytoplasmic protein extract. (F) Transcriptional activity of the proximal human VEGF-A promoter was measured using a luciferase vector with luciferase driven by the VEGF-A promoter (spanning base pairs −2018 to +50 bp relative to the transcription start site). Transfected 293 cells were infected with 3 pfu per cell of either wild-type HSV-1 strains; McKrae, KOS, or SC16 or with 3 pfu per cell HSV-2 and assayed for luciferase activity at the indicated time PI. Luciferase activity was normalized to the activity of uninfected 293 cells transfected with a promoterless luciferase vector, pGL3 (** p<0.01). All figures are representative figures from individual experiments with an n = 3/group for A–D. Extracts from 3 culture wells per time point were pooled for E. Bars denote mean VEGF-A pg per μg of cytoplasmic protein ± SEM.
From: Wuest T, Zheng M, Efstathiou S, Halford WP, Carr DJJ (2011)
The Herpes Simplex Virus-1 Transactivator Infected Cell Protein-4 Drives VEGF-A Dependent Neovascularization.
PLoS Pathog 7(10): e1002278.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Sheep anti Green Fluorescent Protein antibody used for the identification of GFP tagged VEGF-A expressing cells by immunofluorescence.
Image caption:
The HSV-1 transactivator ICP4 is required for VEGF-A expression.
Human 293 cells were plated and infected with 3 pfu per cell HSV-1 strain McKrae. Fold induction of VEGF-A transcript expression was determined via geometric means of fold induction of VEGF-A transcript relative to the housekeeping genes β-actin, TBP and PPIA. (A) Inhibition of protein synthesis with cycloheximide (100 μg/mL) blocked upregulation of VEGF-A following HSV-1 infection while (B) inhibition of viral DNA synthesis with acyclovir (200 μM) had no significant effect. (C) ICP4 dependence was tested using human 293 cells infected with 3 pfu per cell of wild type HSV-1 KOS or KOS-derived mutants lacking ICP4 (ICP4− null virus) or the HSV-1 origin binding protein (OBP− null virus). VEGF-A transcript was measured via real time RT-PCR relative to β-actin, TBP and PPIA. (*p<0.05 relative to uninfected control cells). (D) Human 293 cells transfected with pVA8855 or the promoterless luciferase control plasmid, pGL3. Luciferase activity was measured at 12 hours PI with HSV-1 KOS, ICP4− null, or OBP− null virus. (E) VEGF-A mRNA levels in human 293 cells infected with 3 pfu per cell HSV-1KOS or an HSV-1 ICP0− null virus that fails to make the IE co-transactivator protein, ICP0; both viruses significantly up-regulated VEGF-A mRNA levels. (F) Mice lacking type I interferon responses due to deficiency in the type I interferon receptor (CD118−/−) were infected with HSV-1 KOS or HSV-1 ICP0− null virus. Corneas were harvested at 24 hours PI with 105 pfu of HSV-1 and analyzed for VEGF-A levels by cytokine bead array, expressed as pg of VEGF-A per mg of cornea wet mass. VEGF-A was induced by inoculation with HSV-1 KOS or HSV-1 ICP0− null virus (** p<0.01 relative to uninfected, scarified controls). (G) Reporter mice expressing GFP under the human VEGF-A promoter were analyzed for HSV-1 antigen (red) and GFP (green) along with DAPI (blue) at 12 hours PI with either HSV-1 KOS or HSV-1 ICP4- virus. A and B are representative figures of two experiments, n = 3/group/experiment. Fold induction values were normalized to VEGF-A levels in uninfected, vehicle-treated controls. Panels C and D are representative of two experiments, n = 3/group/experiment. Panel E is a summary of two experiments, n = 6/group. Panel F is representative of two experiments with an n = 3/group/experiment. Bars denote mean ± SEM. (** p<0.01, *<0.05, NSD = non-significant difference).
From: Wuest T, Zheng M, Efstathiou S, Halford WP, Carr DJJ (2011)
The Herpes Simplex Virus-1 Transactivator Infected Cell Protein-4 Drives VEGF-A Dependent Neovascularization.
PLoS Pathog 7(10): e1002278.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Goat anti Green Fluorescent Protein
- Product Type
- Polyclonal Antibody
- Isotype
- Polyclonal IgG
- Specificity
- Green Fluorescent Protein
Antibody Quality and Performance Guarantee
| Goat anti green fluorescent protein antibody recognizes green fluorescent protein (GFP), a 27kD protein which is derived from the jellyfish Aequorea victoria. GFP fluoresces green (509nm) when excited by blue light (395nm) and is commonly used as a marker of gene expression. The antibody recognises wild type GFP, but will also detect recombinant and enhanced forms of GFP. |
- Product Form
- Purified IgG - liquid
- Antiserum Preparation
- Antisera to green fluorescent protein (GFP) were raised by repeated immunisations of goat with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
- Immunogen
- Fusion protein corresponding to full length GFP derived from Aequorea victoria
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
1/10,000 | 1/30,000 |
| Immunofluorescence | |
||
| Western Blotting | |
1/400 | 1/10,000 |
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Rabbit anti Goat IgG (Fc):FITC | STAR122F | F | 1 mg |
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| Description | Rabbit anti Goat IgG (Fc):FITC | ||||||
| Rabbit anti Goat IgG (Fc):HRP | STAR122P | C E WB | 1 mg |
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| Description | Rabbit anti Goat IgG (Fc):HRP | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| Rabbit anti Red Fluorescent Protein | AHP2987 | WB | 50 µg |
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| Description | Rabbit anti Red Fluorescent Protein | ||||||
References for Green Fluorescent Protein antibody
-
Wen, J, & Brogna, S. (2010) Splicing-dependent NMD does not require the EJC in Schizosaccharomyces pombe.
EMBO J. 29: 1537-51. -
Wuest, T. et al. (2011) The Herpes Simplex Virus-1 Transactivator Infected Cell Protein-4 Drives VEGF-A Dependent Neovascularization.
PLoS Pathog. 7: e1002278. -
Al-Jubran, K. et al. (2013) Visualization of the joining of ribosomal subunits reveals the presence of 80S ribosomes in the nucleus.
RNA. 19: 1669-83. -
Lampron, A. et al. (2013) Migration of bone marrow-derived cells into the central nervous system in models of neurodegeneration.
J Comp Neurol. 521 (17): 3863-76. -
Nicol, M.Q. et al. (2019) Lack of IFNγ signaling attenuates spread of influenza A virus in vivo and leads to reduced pathogenesis.
Virology. 526: 155-164. -
Berger, S.L. et al. (2018) Localized Myosin II Activity Regulates Assembly and Plasticity of the Axon Initial Segment.
Neuron. 97 (3): 555-570.e6.
- RRID
- AB_566990
- UniProt
- P42212
- GO Terms
- GO:0006091 generation of precursor metabolites and energy
- GO:0008218 bioluminescence
- GO:0018298 protein-chromophore linkage
AHP975
152293
164647
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