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Invitrogen
CD24 Monoclonal Antibody (M1/69), NovaFluor™ Yellow 660, eBioscience™
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Application
FIGURE: 1 / 4
CD24 Antibody (M014T02Y04-A) in Flow
BALB/c mouse splenocytes were unstained (left) or stained with 0.8 µg of CD24 Monoclonal Antibody, NovaFluor Yellow 660 (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0193-82). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora.
Product Details
M014T02Y04-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
M1/69
Conjugate
Excitation/Emission Max
550/664 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3098994 (AB_2896809)
Product Specific Information
Description
The M1/69 monoclonal antibody reacts with the mouse CD24 molecule, also known as Heat Stable Antigen (HSA). The expression of CD24 has been used to resolve stages of B lymphopoiesis in mouse bone marrow.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The M1/69 antibody has been tested by flow cytometric analysis of mouse splenocytes and thymocytes. This can be used at less than or equal to 0.8 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Our internal testing shows that NovaFluor Yellow 660 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
Excitation: 550 nm; Emission: 664 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD24 is a 35-50 kDa glycosylphosphatidylinositol (GPI)-anchored glycoprotein expressed on a variety of cell types, including erythrocytes, thymocytes, peripheral lymphocytes, and cells of the myeloid lineage. It is variably glycosylated, resulting in heterogeneity in molecular mass across different cell lineages, which can affect antibody staining levels on lymphocyte populations. In the context of B-cell development, CD24 is expressed from the pro-B-cell stage in the bone marrow through to mature, surface Ig-positive B cells, with very low or negative expression on plasma cells. It is also present on the majority of B-lineage acute lymphoblastic leukemias, B-cell chronic lymphocytic leukemias (CCLs), and B-cell non-Hodgkin's lymphomas. CD24 is thought to play a role in regulating B-cell proliferation and maturation, as well as in controlling autoimmunity. Additionally, CD24 has been reported to interact with P-selectin (CD62P), indicating a potential role in cell adhesion and signaling. CD24 is a valuable marker for studying B-cell development, leukemia, lymphoma, and autoimmune regulation, as well as its interactions with other molecules like P-selectin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: CD24; cluster of differentiation 24; FLJ22950; FLJ43543; heat stable antigen; HSA; Ly-52; Lymphocyte antigen 52; M1/69-J11D heat stable antigen; MGC75043; Nectadrin; R13-Ag; Signal transducer CD24; X62 heat stable antigen
Gene Aliases: Cd24; Cd24a; HSA; Ly-52; nectadrin
UniProt ID: (Mouse) P24807
Entrez Gene ID: (Mouse) 12484
Performance Guarantee
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