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Invitrogen
CD22 Monoclonal Antibody (eBio4KB128 (4KB128)), NovaFluor™ Blue 690, eBioscience™
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FIGURE: 1 / 2
CD22 Antibody (H053T03B09-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD22 Monoclonal Antibody, NovaFluor Blue 690 (right). All cells were co-stained with CD19 Monoclonal Antibody, eFluor 450 (Product # 48-0199-42). 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 and unmixed with autofluorescence extraction.
Product Details
H053T03B09-A
Product Specifications
Species Reactivity
Human
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
eBio4KB128 (4KB128)
Conjugate
Excitation/Emission Max
672/688 nm
View spectra
Form
Liquid
Concentration
0.2 µg/Test
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2, with BSA
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
AB_3251402
Product Specific Information
Description
The eBio4KB128 monoclonal antibody recognizes human CD22 (Siglec-2), which is a member of the siglec subgroup of the Ig superfamily.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This eBio4KB128 (4KB128) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.2 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.
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
Excitation: 494 nm; Emission: 688 nm; Laser: 488 nm (Blue) Laser
Target Information
CD22, also known as BL-CAM, is a type I transmembrane glycoprotein composed of two polypeptide chains, CD22alpha and CD22beta, with molecular weights of 130 and 140 kDa, respectively. These chains are produced by alternative splicing of the CD22 gene. CD22 is prominently expressed on mature B cells and B cell lymphomas, including hairy cell leukemia, diffuse large B-cell lymphoma, and nodular lymphocyte predominance Hodgkin's lymphoma, but is negative in classical Hodgkin's lymphoma. The extracellular portion of CD22 contains seven Ig-like domains that preferentially bind alpha2,6-linked sialic acid moieties found on epithelial, endothelial, B, and T cells. This binding can be masked by cis interactions with sialic acids on the same cell surface. CD22 expression is limited to late stages of B-cell differentiation, making it useful for phenotyping mature leukemias. Intracellularly, CD22 features six tyrosine residues within immunotyrosine-based inhibitory motifs (ITIM) and activation-like motifs. These residues are phosphorylated upon B-cell receptor engagement, allowing CD22 to regulate B-cell receptor signaling. CD22 participates in positive regulation through interactions with Src family tyrosine kinases and acts as an inhibitory receptor by recruiting cytoplasmic phosphatases via SH2 domains, which block signal transduction through dephosphorylation of signaling molecules. CD22's role in both positive and negative regulation of B-cell signaling, along with its specific expression pattern, makes it a valuable marker for antibody customers interested in B-cell-related research and diagnostics.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
How to use the Panel Builder
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
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Bioinformatics
Protein Aliases: B-cell receptor CD22; B-lymphocyte cell adhesion molecule; B-lymphocyte cell adhesion molecule (BL-CAM); BL-CAM; CD22; CD22 antigen; FLJ22814; Lectin 2; Leu-14; MGC130020; sialic acid binding Ig-like lectin 2; Sialic acid-binding Ig-like lectin 2; Sialic acid-binding Ig-like lectin 2 (Siglec-2); Siglec-2; T-cell surface antigen Leu-14
Gene Aliases: CD22; SIGLEC-2; SIGLEC2
UniProt ID: (Human) P20273
Entrez Gene ID: (Human) 933
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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