Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [2]
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- Product number
- 78-0279-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD27 Monoclonal Antibody (O323), Super Bright™ 780, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The O323 monoclonal antibody reacts with human CD27, a lymphocyte-specific member of the TNFR superfamily. CD27 is expressed by a subset of thymocytes and virtually all mature T cells and is upregulated upon T-cell stimulation. CD27 binds to CD70, and through this interaction, plays an important role in T cell-B cell interaction. Applications Reported: This O323 antibody has been reported for use in flow cytometric analysis. Applications Tested: This O323 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 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. Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497). Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- O323
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Expression of CD27 and CD28 on γδ T cells from the peripheral blood of patients with allergic rhinitis.
More than one antibody of individual B cells revealed by single-cell immune profiling.
Wang Q, Sun Q, Chen Q, Li H, Liu D
Experimental and therapeutic medicine 2020 Dec;20(6):224
Experimental and therapeutic medicine 2020 Dec;20(6):224
More than one antibody of individual B cells revealed by single-cell immune profiling.
Shi Z, Zhang Q, Yan H, Yang Y, Wang P, Zhang Y, Deng Z, Yu M, Zhou W, Wang Q, Yang X, Mo X, Zhang C, Huang J, Dai H, Sun B, Zhao Y, Zhang L, Yang YG, Qiu X
Cell discovery 2019;5:64
Cell discovery 2019;5:64
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells cells were stained with CD19 Monoclonal Antibody, FITC (Product # 11-0119-82) and Mouse IgG1 kappa Isotype Control, Super Bright 780 (Product # 78-4714-82) (left) or CD27 Monoclonal Antibody, Super Bright 780 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 4 Sanger sequencing validation of multiple V(D)J recombination patterns in single B cells. a Sketching diagram of the single-cell sequencing procedure. We isolated peripheral blood from healthy donors and separated peripheral blood mononuclear cells (PBMCs). Naive B cells (CD19 + CD38 +/- CD10 - CD27 - ), memory B cells (CD19 + CD38 +/- CD10 - CD27 + ), and plasma cells (CD19 + CD38 +++ CD10 - ) were sorted by FACS. The total mRNA in single B cells was reverse transcribed, and the Ig heavy chain and light chain were amplified by multiplex PCR. FACS, fluorescence-activated cell sorting. b Proportions of cells expressing VDJ recombination patterns in single B cells from donor 7, donor 8, and donor 9. c - e Distribution and frequency of IGHV ( c ), IGHD ( d ), and IGHJ ( e ) segments in Ig germline genes in a single cell. f Proportions of single naive B cells, plasma cells, and memory B cells expressing one, two, or three V H DJ H segments. g , h Proportions of single B cells expressing one, two, or more V kappa J kappa segments ( g ) and V lamda J lamda segments ( h ). i Counts of single B cells expressing only Igkappa or Iglamda or expressing both Igkappa and Iglamda. j , k Proportions of naive B cells, plasma cells, and memory B cells expressing one, two, three, or four Ig classes
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 Expression pattern of CD27 and CD28 on gammadelta T cells in AR. (A) Representative flow cytometry plots of the expression of CD27 and CD28 on gammadelta T cells among PBMCs from HC (n=12) and AR (n=14) subjects. (B-G) Quantification of the percentage of (B) CD27+, (C) CD28+, (D) CD27 + CD28 + , (E) CD27 + CD28 - , (F) CD27 - CD28 + and (G) CD27 - CD28 - gammadelta + T cells in PBMCs. Values are expressed as the mean +- standard deviation. AR, allergic rhinitis; HC, healthy controls; PBMCs, peripheral blood mononuclear cells.