IgG1 Negative Control conjugated to FITC
| Code | Size | Price |
|---|
| GM-4992 | 2ml (100 Tests) | £138.00 |
Quantity:
Prices exclude any Taxes / VAT
Overview
Antibody Isotype: IgG1
Antibody Clonality: Monoclonal
Antibody Clone: VI-AP
Regulatory Status: RUO
Target Species: Mouse
Application: Flow Cytometry
Storage:
Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protec
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Further Information
Applications Description:
Direct Immunofluorescence (Staining Procedure) Nordic-MUbio fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations.
Proposed staining procedure for whole blood in short:
- For each sample add 50 ?l of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ?l of the appropriate Nordic-MUbio monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4°C or at room temperature in the dark
- Add 100 ?l NM-LYSE (Cat.No. GAS-003) to each tube and incubate for 10 minutes at room temperature
- Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature
- Centrifuge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid
- Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours
For ?No-Wash? protocol please refer to www.nordicmubio.com
Proposed staining procedure for MNC in short:
- Carefully add 20 ?l antibody conjugate and 50-100 ?l MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8°C or at room temperature
- Centrifuge tubes for 5 minutes at 300 g
- Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g
- Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours
Proposed staining procedure for whole blood in short:
- For each sample add 50 ?l of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ?l of the appropriate Nordic-MUbio monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4°C or at room temperature in the dark
- Add 100 ?l NM-LYSE (Cat.No. GAS-003) to each tube and incubate for 10 minutes at room temperature
- Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature
- Centrifuge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid
- Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours
For ?No-Wash? protocol please refer to www.nordicmubio.com
Proposed staining procedure for MNC in short:
- Carefully add 20 ?l antibody conjugate and 50-100 ?l MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8°C or at room temperature
- Centrifuge tubes for 5 minutes at 300 g
- Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g
- Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours
Background:
This ready to use negative control reagent contains purified, fluorescein or phycoerythrin conjugated mouse immunoglobulin molecules of IgG1 isotype, which have been selected on the basis of their binding characteristics: no specific binding to human cell surface or intracellular antigens, same low range of nonspecific binding to human leukocytes as other Nordic-MUbio Reagents.
This isotype control IgG1 is suitable as a Negative Control to be used in combination with Nordic-MUbio reagents for the:
- Enumeration of Myeloid Cells
- Analysis of Myeloid Differentiation Stage
- Enumeration of B-cells and Precursors
- Enumeration of T-cells and Precursors
- Analysis of Leukemia Cells
- Analysis of Immunodeficiency States
The negative control reagent permits to estimate the degree of non-specific binding of isotype matched immunologbulins to leukocytes via e.g. Fc-receptors. It enables the expert to set flow cytometric parameters accordingly.
Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation.
This isotype control IgG1 is suitable as a Negative Control to be used in combination with Nordic-MUbio reagents for the:
- Enumeration of Myeloid Cells
- Analysis of Myeloid Differentiation Stage
- Enumeration of B-cells and Precursors
- Enumeration of T-cells and Precursors
- Analysis of Leukemia Cells
- Analysis of Immunodeficiency States
The negative control reagent permits to estimate the degree of non-specific binding of isotype matched immunologbulins to leukocytes via e.g. Fc-receptors. It enables the expert to set flow cytometric parameters accordingly.
Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation.
Caution:
For professional users only.
This reagent contains sodium azide. To avoid the development
of hazardous conditions, reagents containing azide should be
diluted in running water prior to be discarded. Similar to the work
with other biological products, proper handling procedures are
recommended.
This reagent contains sodium azide. To avoid the development
of hazardous conditions, reagents containing azide should be
diluted in running water prior to be discarded. Similar to the work
with other biological products, proper handling procedures are
recommended.
Formulation:
PBS pH 7.2, 1% BSA, 0.05% NaN3
Label:
FITC
Product:
2ml of FITC-conjugated VI-AP in PBS pH 7.2, 1% BSA, and 0.05% NaN3, approximately 100 tests.
Product Form:
FITC
Specificity:
The clone VI-AP reacts with calf intestine alkaline phosphatase and does not show cross-reactivity with human proteins.
The sensitivity of VI-AP mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb
dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI
(mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 ?l of leukocytes containing 10^7 cells/ml are stained with 20 ?l mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
The sensitivity of VI-AP mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb
dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI
(mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 ?l of leukocytes containing 10^7 cells/ml are stained with 20 ?l mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.