Human Rac1 ELISA Kit (A310412)

1 Citation

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This product is discontinued

Human Rac1 ELISA Kit (A310412) has been discontinued and is no longer available.

View all Rac1 ELISA Kits.

Specifications
Components

Name

Human Rac1 ELISA Kit

Description

Human Rac1 ELISA Kit is a 90 minute sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Rac1 in serum, plasma, and other biological fluids.

Assay Type

Sandwich (quantitative)

Principle of Assay

Human Rac1 ELISA Kit (A310412) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human Rac1 in serum, plasma or other biological fluids. An antibody specific for Rac1 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Rac1 present in each sample is bound to the wells by the immobilized antibody. Biotinylated Anti-Rac1 Antibody, which also binds the Rac1 present in each sample, and Streptavidin-HRP, which binds the Biotinylated Anti-Rac1 Antibody, are added and the microtiter plate is incubated. Following incubation, unbound Biotinylated Anti-Rac1 Antibody and unbound Streptavidin-HRP are removed by washing, and two substrate solutions are added to the wells. Color develops in proportion to the amount of Rac1 captured in each well. The color development is stopped by addition of stop solution which changes the color from blue to yellow and the intensity of the color is then measured. The concentration of Rac1 in the samples can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

Platform

Pre-coated Microplate (12 x 8 well strips)

Detection Type

Colorimetric

Instrument

Colorimetric Microplate Reader

Sample Type

Serum, plasma or other biological fluids.

Sensitivity

0.093 ng/ml

Product Range

2-32 ng/ml

Assay Time

1h 30m

Reactivity

Human

Precision

Intra-assay CV: 6.3%, Inter-assay CV: < 10%

Storage

Store at +4°C. Do not use past expiration date!

Synonyms

Cell migration-inducing gene 5 protein, p21-Rac1, Ras-like protein TC25, Ras-related C3 botulinum toxin substrate 1, TC25

Product Note

Information online is representative. Always refer to the Product Manual inside the kit.

Disclaimer

This product is for research use only. It is not intended for diagnostic or therapeutic use.

Kit Components

Item	Quantity	Storage
Pre-Coated 96 Well Microplate	12 x 8 Well Strips	+4°C
Standard Solution	500µl	+4°C
Standard Diluent	3ml	+4°C
Biotinylated Detection Antibody	1ml	+4°C
Streptavidin-HRP	6ml	+4°C
Wash Buffer (25X)	20ml	+4°C
Substrate Solution A	6ml	+4°C
Substrate Solution B	6ml	+4°C
Stop Solution	6ml	+4°C
Plate Sealers	5 Adhesive Strips	-
Foil Pouch	1 Zip-Sealed Pouch	-

Required Materials

The following materials are not included in the kit, but are required to run this assay:

Microplate reader capable of measuring absorbance at 450nm ± 10nm.
Squirt bottle, multichannel pipette reservoir, or automated microplate washer.
Graph paper or computer software capable of generating logarithmic functions.
Test tubes and Eppendorf tubes to prepare standards and samples.
Multi- and single-channel pipettes and sterile pipette tips.
Deionized or distilled water.
Absorbent paper towels.
37°C incubator.

Important Notes

Samples cannot be diluted for use with this kit. Owing to the material used to prepare the kit, sample matrix interference may falsely depress the specificity and accuracy of the assay.
Sample concentrations should be estimated before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their experiments.
Samples containing Sodium Azide (NaN₃) cannot be used in this assay. NaN₃ inhibits the activity of Horseradish Peroxidase (HRP) which is used in the detection step.
This ELISA kit is intended for the analysis of biological samples and has not been validated for purified/recombinant proteins, industrial, or commercial materials. Performance with such samples cannot be guaranteed, as differences in expression, sequence, tags, folding, post-translational modifications, purity, activity, or buffer composition may affect assay results. For further guidance, please contact our technical support team.

Scientific Validation DataValidation Data (1)

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Standard Curve - Human Rac1 ELISA Kit (A310412)

Representative standard curve for Human Rac1 ELISA Kit (A310412).

Product Citations (1)

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Interferon gamma applied ex vivo restores function to neutrophils from critically ill patients

This publication cites Human Rac1 ELISA Kit (A310412).

Article Abstract

Introduction: Critically ill patients commonly develop acquired neutrophil dysfunction, which increases susceptibility to intensive care unit-acquired infection (ICU-AI). This study aimed to assess whether interferon gamma (IFN-?) can restore function in dysfunctional neutrophils from critically ill patients and to uncover potential underlying mechanisms.

Methods: This was an observational cohort study. Neutrophils were isolated from whole blood donated by critically ill patients (n=31) in four separate teaching hospital intensive care units (ICUs). Neutrophils were subsequently treated with recombinant human IFN-? or vehicle for 1 hour following either Fc gamma receptor (Fc?R) blockade, selective inhibition of the gamma isoform of phosphoinositide 3-kinase (PI3K-?) or vehicle control for 30 min. Neutrophil phagocytosis, bacterial killing, superoxide generation, phagocytic receptor expression and small Rho GTPase activity were assessed. Neutrophil dysfunction was defined as <50% of cells ingesting 2 or more zymosan particles in a phagocytosis assay.

Results: IFN-? significantly improved phagocytosis (control 36.5%, IFN-? 56.0%), bacterial killing (control 31.6%, IFN-? 82.1%) and superoxide generation (2.8-fold increase relative to control) in dysfunctional neutrophils. IFN-? also increased the activity of the small GTPases, Rac and Cdc42 (2.4-fold and 1.5-fold increase relative to control, respectively) in dysfunctional neutrophils. Selective inhibition of PI3K-? prevented the IFN-?-mediated improvement of phagocytosis (IFN-? 62.5%, with inhibitor 27.9%), bacterial killing (IFN-? 82.1%, with inhibitor 30.5%) and superoxide generation (IFN-? 2.8-fold change relative to control, 0.7 with inhibitor). The IFN-?-mediated improvement of bacterial killing in dysfunctional neutrophils was also prevented by Fc?R blockade (IFN-? 82.1%, Fc?R inhibition 28.7%).

Conclusions: In critically ill patients with known acquired neutrophil dysfunction, ex vivo application of IFN-? consistently improved a range of neutrophil effector functions.

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