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GFP Antikörper (Green Fluorescent Protein)

Details für Produkt anti-GFP Antikörper Nr. ABIN398304, Anbieter: Anmelden zum Anzeigen
Antigen
  • green fluorescent protein
  • gfp
Reaktivität
Aequorea victoria
648
5
4
3
2
2
2
1
Wirt
Ratte
327
172
87
44
16
9
7
2
Klonalität (Klon)
Monoklonal ()
Konjugat
Dieser GFP Antikörper ist unkonjugiert
34
33
29
15
13
9
9
8
7
7
6
6
6
5
5
5
5
5
5
4
4
3
3
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
Applikation
ELISA, Western Blotting (WB)
536
351
200
142
138
134
76
70
58
26
13
13
10
9
8
6
6
4
4
3
3
3
3
3
2
2
2
1
1
1
1
1
Optionen
Hersteller
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Bilder

'Independent Validation' Siegel
Chargennummer 100316
Validierte Anwendung Western Blotting
Positivkontrolle 293FT cells transduced with eGFP virus
Negativkontrolle untransduced 293FT cells
Bewertung A major band was observed at the correct molecular weight in the two positive (eGFP-transduced) cell lysates. No major bands were observed in the negative control (untransduced cells). Some additional faint bands of lower molecular weight were also observed in the positive controls, this may be because the antibody was used at 1:500 instead of the recommended 1:1000 dilution. Microscope images of transduced cells are provided to demonstrate successful eGFP-transduction of cells.
Primärantikörper
Sekundärantikörper
Kontrollen
  • Positive control: 293FT cells transduced with eGFP virus
  • Negative control: untransduced 293FT cells
Protokoll
  • 20 µg of total protein from prepared cell lysates was run on a 4-12% SDS-PAGE gel.
  • The gel was run at 180 V for 40 minutes and transferred to a PVDF membrane using a wet-transfer apparatus.
  • The membrane was rinsed in TBST for 10 min in TBST.
  • The membrane was blocked in 3% BSA for 30 min.
  • The membrane was incubated in 1:500 primary antibody diluted in 3% BSA at 4°C overnight.
  • The membrane was washed 3 times for 10 min in TBST.
  • The membrane was incubated in secondary antibody at 1:10000 for 1 hr at RT.
  • The membrane was washed 3 times for 10 min in TBST.
  • The membrane was imaged using a fluorescent imaging system at 679nm⁄702nm.
Anmerkungen Fluorescent microscopy images are included to confirm eGFP transduction of 293FT cells.
Validierungsbilder
Western Blotting validation image for anti-Green Fluorescent Protein (GFP) antibody (ABIN398304) Figure 1: A. Western blot of lysates from untransduced 293FT cells (Lane 2) and 293FT...
Verwendungszweck For biochemical analysis of GFP-tagged fusion proteins.
Klon 3H9
Isotyp IgG2a
Spezifität 3H9 efficiently recognizes green fluorescent proteins from Aequorea victoria such as eGFP, wtGFP, YFP, or CFP, but no red fluorescent proteins.
Produktmerkmale Fluorescent proteins (FP) are powerful tools to study protein localization and dynamics in living cells in particular the prominent green fluorescent protein (GFP) from Aequorea victoria.
Here we offer a rat monoclonal antibody (MAb) against various GFPs, designated as 3H9. The 3H9 MAb is characterized by high affinity and specificity. It is suitable for immunoblotting and ELISA.
With this versatile antibody GFP and fusions thereof become attractive tools for biochemical applications like identification of interacting proteins or Western Blotting.
Reinigung Hybridoma supernatant. The Hybridoma was cultured in protein free medium. Tissue culture supernatant was concentrated by centrifugation using a 30 kDa exclusion membrane to an antibody concentration of 1 mg/ml.
Hintergrund The first green fluorescent protein (GFP) was isolated from Aequorea victoria and has a molecular weight of 27 kDa. Genetic mutations of GFP resulted in fluorescent proteins with different excitation and emission spectra e.g. eGFP, YFP, CFP. Fluorescent proteins became attractive tools for a variety of applications.
Forschungsgebiet Tags/Labels
Applikationshinweise Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 31 kDa (the predicted molecular weight of GFP).
Kommentare

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Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 mg/mL
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Do not freeze.
Lagerung 4 °C
Haltbarkeit 12 months
Bilder des Herstellers
Western Blotting (WB) image for anti-GFP Antikörper (Green Fluorescent Protein) (ABIN398304) Western Blot analysis using anti-GFP antibody 3H9. Primary antibody: 0.2 ug of GFP an...
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Reiner, Hoefle, Hückelhoven: "A barley SKP1-like protein controls abundance of the susceptibility factor RACB and influences the interaction of barley with the barley powdery mildew fungus." in: Molecular plant pathology, 2015 (PubMed).

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Qin, Wolf, Liu, Link, Smets, Mastra, Forné, Pichler, Hörl, Fellinger, Spada, Bonapace, Imhof, Harz, Leonhardt: "DNA methylation requires a DNMT1 ubiquitin interacting motif (UIM) and histone ubiquitination." in: Cell research, Vol. 25, Issue 8, pp. 911-29, 2015 (PubMed).

Traenkle, Emele, Anton, Poetz, Haeussler, Maier, Kaiser, Scholz, Nueske, Buchfellner, Romer, Rothbauer: "Monitoring interactions and dynamics of endogenous beta-catenin with intracellular nanobodies in living cells." in: Molecular & cellular proteomics : MCP, Vol. 14, Issue 3, pp. 707-23, 2015 (PubMed).

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Galili, Dylla, Lüdke, Friedrich, Yamagata, Wong, Ho, Szyszka, Tanimoto: "Converging circuits mediate temperature and shock aversive olfactory conditioning in Drosophila." in: Current biology : CB, Vol. 24, Issue 15, pp. 1712-22, 2014 (PubMed).

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