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Application
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CD45 Antibody (M005T02Y06-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 0.4 µg of CD45 Monoclonal Antibody, NovaFluor Yellow 700 (right). All cells were co-stained with Ter-119 Monoclonal Antibody, eFluor 450 (Product # 48-5921-82). Total viable cells 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
M005T02Y06-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2b, kappa
Class
Monoclonal
Type
Antibody
Clone
30-F11
Conjugate
Excitation/Emission Max
551/701 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
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_3098941 (AB_2896776)
Product Specific Information
Description
The 30-F11 monoclonal antibody reacts with all isoforms of mouse CD45, also known as Leukocyte Common Antigen (LCA). CD45 is expressed by all hematopoietic cells excluding mature erythrocytes and platelets. The cytoplasmic portion of CD45 has tyrosine phosphatase enzymatic activity and plays an important role in activation of lymphocytes.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
This 30-f11 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This may be used at less than or equal to 0.4 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 700 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: 551 nm; Emission: 701 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD45 (LCA, leukocyte common antigen) is a receptor-type protein tyrosine phosphatase (PTP) ubiquitously expressed in all nucleated hematopoietic cells, comprising approximately 10% of all surface proteins in lymphocytes. CD45 is absent on non-hematopoietic cell lines, normal and malignant, non-hematopoietic tissues. CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45 protein exists as multiple isoforms as a result of alternative splicing, differ in their extracellular domains but share identical transmembrane and cytoplasmic domains. CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells. CD45 isoforms differ in their ability to translocate into the glycosphingolipid-enriched membrane domains and their expression depends on cell type and physiological state of the cell. CD45 has been shown to be an essential regulator of T- and B-cell antigen receptor signaling and suppresses JAK kinases to regulate cytokine receptor signaling. CD45 is also important in promoting cell survival by modulating integrin-mediated signal transduction pathway, DNA fragmentation during apoptosis and inhibition or upregulation of various immunological functions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: CD45; L-CA; Leukocyte common antigen; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; Receptor-type tyrosine-protein phosphatase C; T200; T220 and B220
Gene Aliases: B220; Cd45; CD45R; L-CA; loc; Ly-5; Lyt-4; Ptprc; T200
UniProt ID: (Mouse) P06800
Entrez Gene ID: (Mouse) 19264
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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