Mouse IgG1,IgG2a isotype control (FITC,PE)
Quick Overview for Mouse IgG1,IgG2a isotype control (FITC,PE) (ABIN1741586)
Target
Conjugate
Application
Clone
Isotype
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Host
- Mouse
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Purpose
- This product is optimised for use with FIX&PERM®.
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Specificity
- The monoclonal antibodies have no known specificity for human leukocytes.It is recommended to use similar amounts of test antibody and isotype matched COMBI-REAGENT Negative Control antibody in each experiment.
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Characteristics
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Mildly fixes cells, preserving their flow cytometric scatter characteristics
Allows simultaneous characterisation of both intracellular and cell surface markers
Rapid technique - whole procedure can be carried out in less than one hour, ready
for immediate analysis or storage for 24 hours
Stringent QC procedures - the quality of each lot is determined using well-defined
blood samples and subsequent comparison of scatter characteristics of obtained
leukocyte populations, ensuring consistent and reliable results lot after lot
A range of intracellular antibodies with optimised protocols for use with FIX&PERM®
FIX&PERM® is a simple procedure making use of two reagents. Reagent A gently fixes cells, while Reagent B permeabilizes them. The specific formulations reduce background and allow simultaneous addition of permeabilization medium and fluorochrome labelled antibodies, allowing staining of intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins, etc. -
Purification
- Purified by Affinity Chromatography
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Components
- COMBI Surface: IgG2a Negative Control (FITC) and IgG1 Negative Control (PE)
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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 µL of EDTA anti-coagulated blood to a 3-5 mL tube - Add 20 µL of the appropriate monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4 °C or at room temperature in the dark - Add 100 µL NM-LYSE (ABIN1741575) 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 Proposed staining procedure for MNC in short: - Carefully add 20 µL antibody conjugate and 50-100 µL MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-4 °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 2-4 °C in the dark. Analyze fixed cells within 24 hours
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Comment
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Each staining performed with specific monoclonal antibodies should be paralleled by a staining with an appropriate istotype matched control antibody, in order to be able to control for non-specific binding. The COMBI-REAGENT-Negative Control 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. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained, which cannot be attributed to differences in laboratory procedures, please contact us.
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Assay Procedure
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Procedure: For each sample to be analysed use 50 µL of whole blood, bone marrow or mononuclear cell suspension in a 5 mL tube
Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature)
Incubate for 15 minutes at room temperature
Add 5 mL phosphate buffered saline and centrifuge cells for 5 minutes at 300 g
Remove supernatant and add to cell pellet 100 µL Reagent B (Permeabilization
Medium) and 20 µL of the appropriate monoclonal antibody conjugate
Vortex at low speed for 1-2 seconds
Incubate for 15 minutes at room temperature
Wash cells with phosphate buffered saline as described above
Remove supernatant and re-suspend cells in sheath fluid for immediate analysis or re-suspend cells in 0.5 mL 1.0 % formaldehyde and store at 2-8 °C in the dark
Analyse fixed cells within 24 hours -
Restrictions
- For Research Use only
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Buffer
- PBS pH 7.2, 1 % BSA, 0.05 % sodium azide
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Preservative
- Sodium azide
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Precaution of Use
- 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.
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Handling Advice
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Do not freeze and protect 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. -
Storage
- 4 °C
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Storage Comment
- These reagents should be stored at 2-8 °C
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- IgG1,IgG2a
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Sub Type
- Cocktail
Target
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