Protein G (20nm Gold) (ab270602)
Overview
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Product name
Protein G (20nm Gold) -
General notes
Protein G Gold Conjugate (20nm, 10 OD) (ab270602) is manufactured using our Gold Nanoparticles which have a protective coating that can withstand the most extreme conditions (e.g. 2.5 M NaOH at 70°C for >1 hour). The irreversible covalent attachment does not allow the desorption of the Protein A, which is a major problem associated with gold conjugates when developed using passive adsorption techniques.
To exchange the conjugate into a specific buffer for your assay or test, centrifuge the conjugate in a microfuge at 9,000 x g for 20 minutes. Carefully remove the supernatant and add your preferred buffer. It is important to avoid substances that have a very high affinity for gold (e.g. thiols).
The maximum absorbance for Protein G Gold Conjugate (20nm, 10 OD) is at 528 nm. To determine the effective concentration of the conjugate obtained we advise to measure the Absmax of light using an UV-vis spectrophotometer after diluting your sample to an appropriate range for your piece of equipment (e.g. if a 40 nm gold conjugate is at 10 OD and is diluted 1:10 the Abs530nm for a 1 cm light path is expected to be around 1 AU).
Before using Protein G Gold Conjugate (20nm, 10 OD) to detect a specific antibody we recommend that you check the affinity of Protein A for that antibody and subtype (see table below). Rabbit and human IgGs, for example, are good candidates because they bind to Protein A with high affinity.
Species
IgG
Binding strength
Rabbit
IgG
High
Human
IgG
High
Mouse
IgG1
Medium
Mouse
IgG2a
High
Mouse
IgG2b
Medium
Mouse
IgG3
Medium
Mouse
IgM
No binding
Goat
IgG
High
Sheep
IgG
High
Rat
IgG1
Weak/Medium
Rat
IgG2a
High
Rat
IgG2b
Weak/Medium
Rat
IgG2c
Weak/Medium
Protein G Gold Conjugate (20nm, 10 OD) is tested as a detector tool in Lateral Flow Assay but can also be used in electron and light microscopy, immunoblotting, and dot-blot. As Protein G Gold Conjugate (20nm, 10 OD) affinity for antibodies can vary, a titration of the specific antibody used is recommended in order to obtain the optimal performance. We recommend performing the incubation of Protein G Gold Conjugate (20nm, 10 OD) with the antibody in a buffer that has a pH between 5 and 7.5 for at least 10-15 minutes. After this stage do not centrifuge the gold complex.
This product is manufactured by Expedeon, an Abcam company. It was previously called 20nm InnovaCoat® GOLD (10 OD) Protein G Conjugate. Expedeon product code 225-0200 is the same as the 200 µl size of this product and code 225-1000 is the same as the 1 mL size.
Properties
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Form
Liquid -
Storage instructions
Shipped at Room Temperature. Store at +4°C. -
Storage buffer
pH: 7
Constituents: Tris buffered saline, 0.1% Tween -
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