Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- 63-1379-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD137 (4-1BB) Monoclonal Antibody (4B4 (4B4-1)), Super Bright™ 600, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This 4B4 (4B4-1) monoclonal antibody reacts with human CD137 (also known as 4-1BB or TNFRSF9), which is an inducible member of the TNFR family of costimulatory molecules expressed on T cells, natural killer cells, dendritic cells, granulocytes, and mast cells. Involved in recruiting TNFR-associated factors (TRAF) 1 and 2, CD137 signaling plays a role in T cell activation, maintaining the survival of activated and CD8 memory T cells, as well as suppressing myelopoiesis and dendritic cell development. Stimulation of this receptor has also been shown to promote expansion of CD4+CD25+ T regulatory cells i{ex vivoi}. The ligand for CD137, 4-1BBL, is found on activated macrophages, mature B cells, hematopoietic stem cells, and myeloid progenitor cells. Applications Reported: This 4B4 (4B4-1) antibody has been reported for use in flow cytometric analysis. Applications Tested: This 4B4 antibody has been tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at less than or equal to 5 µL (0.5 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. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 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. 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: 600 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
- 4B4 (4B4-1)
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Neoantigens retention in patient derived xenograft models mediates autologous T cells activation in ovarian cancer.
Want MY, Konstorum A, Huang RY, Jain V, Matsueda S, Tsuji T, Lugade A, Odunsi K, Koya R, Battaglia S
Oncoimmunology 2019;8(6):e1586042
Oncoimmunology 2019;8(6):e1586042
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells cells were stimulated for 72 hours with Con A (Product # 00-4978-93) and then stained with CD25 Monoclonal Antibody, PE (Product # 12-0259-42) and Mouse IgG1 kappa Isotype Control, Super Bright 600 (Product # 63-4714-82) (left) or CD137 Monoclonal Antibody, Super Bright 600 (right). Total viable cells were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 10.1080/2162402X.2019.1586042-F0004 Figure 4. Neoantigen recognition by autologous T cells in-vitro . a) Ratio of IFN-gamma production in patient's T cells at different time points upon coculture with mutated (MUT) or WT peptide-stimulated autologous APCs. Y axis indicates the ratio IFN-gamma Mut(pg/ml)/IFN-gamma WT(pg/ml) measured via ELISA. Peptides with a ratio > 1 are highlighted in red boxes and utilized for subsequent experiments. ND = no IFN-gamma production detected. b) Flow cytometry analysis of CD137 + T cells expressed as percent of activated CD8 + T cells after overnight stimulation with 10 um mutated (MUT) or WT peptides. c) Flow plots of CD137 + /CD8 + T cells in response to coculture with APC pulsed with mutated (MUT) or WT peptides. d) Line plot showing the changes in variant allele frequency of the six neoantigenic mutations common across the primary, P0 and P1 tumors. Orange lines represent mutations present in all samples, black dotted mutations indicate those present in P0 and P1 but absent (VAF = 0) in the primary tumor.