Mouse IgG1 isotype control (FITC,PE)

Details for Product No. ABIN1741585, Supplier: Log in to see
Isotype
  • IgG1
  • Igh-4
  • VH7183
  • immunoglobulin heavy constant gamma 1 (G1m marker)
  • Ighg1
Host
Mouse
267
86
22
4
3
3
2
2
1
1
1
1
Conjugate
FITC,PE
45
44
31
27
8
8
7
6
5
5
5
4
4
4
3
3
3
3
3
3
3
3
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
Application
Flow Cytometry (FACS), Isotype Control (IsoC), Negative Control (NC)
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Supplier
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Purpose This product is optimised for use with FIX&PERM®.
Brand FIX&PERM®
Clone VI-AP-VI-AP
Isotype IgG1
Specificity VI-AP reacts with calf intestine alkaline phosphatase and does not show cross-reactivity with human proteins.
Characteristics 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
Components COMBI IC Reagent: IgG Negative Control (FITC) and IgG Negative Control (PE)
Sub Type Cocktail
Research Area Secondary Antibodies
Application Notes Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM- (ABIN1741575) intracellular isotype controls can be easily stained in cell suspensions. - For each sample to be analyzed add 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 COMBI-IC Negative Control 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 resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 mL 1.0 % formaldehyde and store them at 2-8 °C in the dark. Analyze fixed cells within 24 hours.
Comment

This ready to use Negative Control reagent contains a combination of fluorescein and phycoerythrin conjugated mouse immunoglobulin molecules of IgG1 isotype, which have been selected on the basis of their binding characteristics: no specific binding to human intracellular or cell surface antigens, same low range of nonspecific binding to human leukocytes as other COMBI-IC-Reagents. Like all other COMBI-IC-reagents, this reagent should be used in combination with our FIX&PERM- Cell Permeabilization Kit (Cat.No. GAS-002) These isotype control IgG1 are suitable as negative controls to be used in combination with COMBI-IC 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 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.

Assay Procedure

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
Buffer PBS pH 7.2, 1 % BSA, 0.05 % sodium azide
Preservative Sodium azide
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.
Handling Advice 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
Storage Comment These reagents should be stored at 2-8 °C
Product cited in: Knibbs, Craig, Natsuka, Chang, Cameron, Lowe, Stoolman: "The fucosyltransferase FucT-VII regulates E-selectin ligand synthesis in human T cells." in: The Journal of cell biology, Vol. 133, Issue 4, pp. 911-20, 1996 (PubMed).