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GFP antibody (Green Fluorescent Protein)

Details for Product anti-GFP Antibody No. ABIN398304, Supplier: Log in to see
Antigen
Reactivity
Aequorea victoria
547
5
3
2
2
2
2
1
Host
Rat
275
155
73
37
10
7
2
Clonality (Clone)
Monoclonal ()
Conjugate
This GFP antibody is un-conjugated
31
30
23
14
12
6
6
4
4
4
3
3
3
3
3
3
3
3
3
3
3
3
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
ELISA, Western Blotting (WB)
462
288
133
122
115
57
56
35
26
23
13
11
10
9
8
4
3
1
1
1
1
1
1
Supplier
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Available images

'Independent Validation' Badge
Lot Number 100316
Method validated Western Blotting
Positive Control 293FT cells transduced with eGFP virus
Negative Control untransduced 293FT cells
Notes 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.
Primary Antibody
  • Antigen: Green Fluorescent Protein (GFP)
  • Catalog number: ABIN398304
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 100316
Secondary Antibody
Controls
  • Positive control: 293FT cells transduced with eGFP virus
  • Negative control: untransduced 293FT cells
Protocol
  • 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.
Experimental 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...
Purpose For biochemical analysis of GFP-tagged fusion proteins.
Clone 3H9
Isotype IgG2a
Specificity 3H9 efficiently recognizes green fluorescent proteins from Aequorea victoria such as eGFP, wtGFP, YFP, or CFP, but no red fluorescent proteins.
Characteristics 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.
Background 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.
Research Area Tags/Labels
Application Notes Western Blot: Used at a dilution of 1/1000. Detects a band of approximately 31 kDa (the predicted molecular weight of GFP).
Comment

IF: not working

Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze.
Storage 4 °C
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for anti-GFP antibody (Green Fluorescent Protein) (ABIN398304) Western Blot analysis using anti-GFP antibody 3H9. Primary antibody: 0.2 ug of GFP an...
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