NeutrAvidin Protein (TRITC)

Cat. No.: rp188521

GRADE & PURITY Validated, ExactAb™, Ex:557nm, Em:576nm, 1.0 mg/mL

Bioactivity

Binding Biotin

Overview

NeutrAvidin Protein is a specially-prepared form of avidin that decreases background in biotin-binding.
Features include:
• Near-neutral isoelectric point—pI = 6.3, more neutral than native avidin
• Nearly devoid of glycosylation—decreased possibility of lectin binding compared to native avidin
• No RYD recognition sequence—no known off-target binding domains like streptavidin
• Affordable—significantly less expensive than streptavidin
NeutrAvidin Protein is deglycosylated native avidin from egg whites. Removal of the excess carbohydrate by an exclusive process yields a protein with a more neutral isoelectric point and less nonspecific binding properties. Purified NeutrAvidin Protein provides exceptional performance in western blot, ELISA, and IHC applications that require biotin-binding probes. Assay specificity, sensitivity, and signal-to-noise ratios with NeutrAvidin Protein are generally equivalent or better than with the significantly more expensive streptavidin.
Avidin is a glycoprotein found in the egg white and tissues of birds, reptiles, and amphibia. The biotin-binding protein contains four identical subunits having a combined mass of 67,000-68,000 daltons. Removing the glycosylation from avidin yields NeutrAvidin Protein with a mass of 60,000 daltons. Carbohydrate-based lectin binding is reduced to undetectable levels, yet biotin-binding affinity is retained. NeutrAvidin Protein offers the advantages of a neutral pI to minimize nonspecific adsorption, along with lysine residues that remain available for derivatization or other customized conjugation. NeutrAvidin Protein yields the lowest nonspecific binding among the known biotin-binding proteins. The specific activity for biotin binding is approximately 14 µg/mg of protein, which is near the theoretical maximum activity.

Specifications

Product Name

NeutrAvidin Protein (TRITC)

Synonyms

Nuetral Avidin | NeutrAvidin | NeutrAvidin - TRITC Conjugated

Grade

Em:576nm, Ex:557nm, ExactAb™, Validated

Specifications & Purity

ExactAb™, Validated, Ex:557nm, Em:576nm, 1.0 mg/mL

Bioactivity

Binding Biotin

Conjugation

TRITC

Excitation (nm)

Green Laser(532nm)

Emission(nm)

576nm

Ex/Em(nm)

Predicted molecular weight

60kDa

Molecule Type

Protein

Storage and Shipping

Concentration

1.0 mg/mL

Storage

Store at 2-8°C,Protected from light,Do not freeze

Shipped In

Wet ice,Do not freeze

Stability And Storage

The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.

Images

IF: NeutrAvidin Protein (TRITC) (rp188521)

NeutrAvidin Protein (TRITC) (rp188521) - IF
IF analysis of beta Tubulin (red) in Hela cells. The cells were fixed and permeabilized with 100% methanol for 5 minutes, and blocked with 2% BSA for 1 hour at room temperature. Cells were stained with beta Tubulin Mouse mAb (Ab155823) at 0.5 μg/mL in blocking buffer overnight at 4°C, and then incubated with Goat Anti-Mouse IgG H&L (Biotin) (Ab179002) at a dilution of 1/1000 for 1 hour at room temperature in the dark. Then incubated with NeutrAvidin Protein (TRITC) (rp188521) at a dilution of 1/1000 for 1 hour at room temperature in the dark (red). PBS instead of the primary antibody was used as the secondary antibody only control. Cells were counterstained with DAPI (blue). Images were taken on the confocal laser scanning microscope.

Flow Cytometry: NeutrAvidin Protein (TRITC) (rp188521)

NeutrAvidin Protein (TRITC) (rp188521) - Flow Cytometry
Flow cytometry analysis of CD45 expression by Jurkat cells (red) compared to an isotype control (blue). Cells were stained with CD45 Mouse mAb (Biotin) (Ab185413) at 0.05μg/test for 1 hour at 4°C. Then incubated with NeutrAvidin Protein (TRITC) (rp188521) at 0.1μg/test for 30 minutes at 4°C in the dark. Blue - Mouse IgG Isotype Control Antibody (Biotin) (Ab180306). Black - Unlabelled control, cells without incubation with primary antibody.

Application

• Application: Dilution info
• IF/ICC:

  1:1000;

• Flow Cytometry:

  ≤0.1 μg/test

  For flow cytometric staining, the suggested use of this reagent is ≤0.1 μg per million cells in 100 µL staining volume.;