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Invitrogen
CD40 Monoclonal Antibody (5C3), NovaFluor™ Red 685, eBioscience™
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CD40 Antibody (H027T03R02-A) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD40 Monoclonal Antibody, NovaFluor Red 685 (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.
产品信息
H027T03R02-A
产品规格
种属反应
Human
宿主/亚型
Mouse
/ IgG1, kappa
分类
Monoclonal
类型
Antibody
克隆号
5C3
偶联物
激发/发射光谱
637/685 nm
查看光谱
形式
Liquid
浓度
0.6 µg/Test
纯化类型
Affinity chromatography
保存液
PBS, pH 7.2
内含物
0.09% sodium azide
保存条件
4°C, store in dark, DO NOT FREEZE!
RRID
产品详细信息
Description
The 5C3 monoclonal antibody reacts with human CD40, a 45-50 kDa type I transmembrane glycoprotein. 5C3 is reported to be used for activation of B cells in in vitro functional assays.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The 5C3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.6 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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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: 637 nm; Emission: 685 nm; Laser: 633-640 nm (Red) Laser
靶标信息
CD40 is a single-chain glycoprotein and a member of the tumor necrosis factor receptor (TNFR) family, exhibiting significant homology to the Hodgkin's disease-associated antigen, CD30. It is expressed by B lymphocytes, follicular dendritic cells, thymic epithelium, and a subset of peripheral T cells, as well as some epithelial cells, carcinomas, and lymphoid dendritic cells. Notably, CD40 is present on all B cells except plasma cells. CD40 plays a crucial role in regulating B cell development and maturation, inducing immunoglobulin isotype-switching, and protecting B cells from surface Ig-induced apoptosis when combined with other signals such as IL-4. It promotes proliferation and is essential for T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. The interaction between CD40 and its ligand CD154 (gp39) on T cells is vital for T-B cell crosstalk, costimulation, and immune regulation. Adaptor protein TNFR2 interacts with CD40, mediating signal transduction, while the AT-hook transcription factor AKNA is reported to regulate CD40 expression, which may be important for homotypic cell interactions. Additionally, the interaction between CD40 and its ligand is necessary for amyloid-beta-induced microglial activation, suggesting a role in the early pathogenesis of Alzheimer's disease. Two alternatively spliced transcript variants encoding distinct isoforms of CD40 have been identified. Diseases associated with CD40 dysfunction include Type 3 Hyper-IgM immunodeficiency and CD40 ligand deficiency, highlighting its importance in immune and inflammatory responses.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
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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生物信息学
蛋白别名: B cell surface antigen CD40; B cell-associated molecule; B-cell surface antigen CD40; Bp50; CD antigen CD40; CD40; CD40 molecule, TNF receptor superfamily member 5; CD40L receptor; CDw40; Immunoglobulin M; ImmunoglobulinM; MGC9013; sCD40; soluble CD40; Tumor necrosis factor receptor superfamily member 5
基因别名: Bp50; CD40; CDW40; p50; TNFRSF5
UniProt ID: (Human) P25942
Entrez Gene ID: (Human) 958
positive regulation of protein phosphorylation
immune response-regulating cell surface receptor signaling pathway
protein complex assembly
cellular calcium ion homeostasis
inflammatory response
immune response
multicellular organism development
platelet activation
positive regulation of B cell proliferation
response to lipopolysaccharide
positive regulation of interleukin-12 production
tumor necrosis factor-mediated signaling pathway
B cell proliferation
regulation of cell proliferation
positive regulation of tyrosine phosphorylation of Stat1 protein
defense response to protozoan
positive regulation of I-kappaB kinase/NF-kappaB signaling
positive regulation of MAP kinase activity
protein kinase B signaling
positive regulation of GTPase activity
positive regulation of transcription from RNA polymerase II promoter
positive regulation of isotype switching to IgG isotypes
regulation of immune response
regulation of immunoglobulin secretion
positive regulation of NF-kappaB transcription factor activity
defense response to virus
cellular response to lipopolysaccharide
cellular response to mechanical stimulus
positive regulation of protein kinase C signaling
apoptotic signaling pathway
positive regulation of endothelial cell apoptotic process
