GFP anticorps (Green Fluorescent Protein)

Details for Product anti-GFP Antibody No. ABIN398304
Antigène
  • green fluorescent protein
  • gfp
Reactivité
Aequorea victoria
477
10
10
9
9
7
6
1
1
Hôte
Rat
200
175
74
35
16
7
2
2
Clonalité (Clone)
Monoclonal ()
Conjugué
Cet anticorp GFP est non-conjugé
26
23
18
13
10
4
3
3
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
ELISA, Western Blotting (WB)
397
227
121
105
99
55
40
29
25
25
23
15
12
11
8
7
6
4
1
1
1
1
Options
'Independent Validation' signe
Numéro du lot 100316
Application validée Western Blotting
Contrôle positif 293FT cells transduced with eGFP virus
Contrôle négative untransduced 293FT cells
Conclusion 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.
Anticorps primaire
  • Antigen: Green Fluorescent Protein (GFP)
  • Catalog number: ABIN398304
  • Lot number: 100316
Anticorps secondaire
  • Antibody: Alexa Fluor 680 Goat Anti-Rat IgG (H+L)
  • Lot number: N/A
Contrôles
  • Positive control: 293FT cells transduced with eGFP virus
  • Negative control: untransduced 293FT cells
Protocole
  • 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.
Notes Fluorescent microscopy images are included to confirm eGFP transduction of 293FT cells.
Validation Images
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...
Fonction For biochemical analysis of GFP-tagged fusion proteins.
Clone 3H9
Isotype IgG2a
Specificité 3H9 efficiently recognizes green fluorescent proteins from Aequorea victoria such as eGFP, wtGFP, YFP, or CFP, but no red fluorescent proteins.
Attributs du produit 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.
Purification 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.
Plasmids, Primers & others Plasmids, Primers & others GFP products on genomics-online (e.g. as negative or positive controls)
Antigène
Sujet 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.
Indications d'application Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 31 kDa (the predicted molecular weight of GFP).
Commentaires

IF: not working

Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Agent conservateur Sodium azide
Précaution d'utilisation This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation Do not freeze.
Stock 4 °C
Date de péremption 12 months
Images (Fournisseur)
Western Blotting (WB) image for anti-Green Fluorescent Protein (GFP) antibody (ABIN398304) Western Blot analysis using anti-GFP antibody 3H9. Primary antibody: 0.2 ug of GFP an...
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