IgG1 Negative Control
IgG1 Negative Control, conjugated to PE
Reactivity
Mouse
Applications
Immunofluorescence, Immunofluorescence, Flow Cytometry, Functional Assay
Synonyms
IgG1 Negative Control; conjugated to PE; IgG1 Negative Control negative control
Reactivity
Mouse
Isotype
IgG1
Clone Number
VI-AP
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 ul of leukocytes containing 10^7 cells/ml are stained with 20 ul 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 ul of leukocytes containing 10^7 cells/ml are stained with 20 ul 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.
Form/Format
PBS pH 7.2, 1% BSA, 0.05% NaN3
Applicable Applications for IgG1 Negative Control negative control
Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS)
Application Notes
Direct Immunofluorescence (Staining Procedure) 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 ul of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ul of the appropriate monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark
- Add 100 ul NM-LYSE 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 degree C in the dark and analyze within 24 hours
Proposed staining procedure for MNC in short:
- Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8 degree 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 degree C in the dark. Analyze fixed cells within 24 hours
Proposed staining procedure for whole blood in short:
- For each sample add 50 ul of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ul of the appropriate monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark
- Add 100 ul NM-LYSE 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 degree C in the dark and analyze within 24 hours
Proposed staining procedure for MNC in short:
- Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8 degree 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 degree C in the dark. Analyze fixed cells within 24 hours
CE Mark
CE
Conjugation
PE
Preparation and Storage
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 degree C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.
Related Product Information for IgG1 Negative Control negative control
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 Reagents.
This isotype control IgG1 is suitable as a Negative Control to be used in combination with 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 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 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 tests as well as the clinical history of the patient by a certified professional before final interpretation.
Similar Products
Product Notes
The IgG1 Negative Control (Catalog #AAA570403) is a Negative Control and is intended for research purposes only. The product is available for immediate purchase. The IgG1 Negative Control, conjugated to PE reacts with Mouse and may cross-react with other species as described in the data sheet. AAA Biotech's IgG1 Negative Control can be used in a range of immunoassay formats including, but not limited to, Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS). Direct Immunofluorescence (Staining Procedure) 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 ul of EDTA anti-coagulated blood to a 3-5 ml tube - Add 20 ul of the appropriate monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark - Add 100 ul NM-LYSE 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 degree C in the dark and analyze within 24 hours Proposed staining procedure for MNC in short: - Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8 degree 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 degree C in the dark. Analyze fixed cells within 24 hours. Researchers should empirically determine the suitability of the IgG1 Negative Control for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "IgG1 Negative Control, Negative Control" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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