Anti-Mouse 4-1BB Monoclonal Antibodies for Flow Cytometry

PA007486.r2a: Recombinant Anti-mouse CD137 (TNFRSF9 or 4-1BB) Monoclonal Antibody (Clone: 3H3), Rat IgG2a Kappa

Recombinant Rat IgG2a Monoclonal Antibody. 
Clone: 3H3.
Isotype:  Rat IgG2a Kappa.
Source: The anti-mouse CD137 (TNFRSF9 or 4-1BB) monoclonal antibody (clone: 3H3) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant 3H3 antibody specifically binds to mouse 4-1BB protein.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

PA007486.h1Fs: Recombinant Anti-mouse CD137 (TNFRSF9 or 4-1BB) Monoclonal Antibody (Clone: 3H3), Human IgG1 Fc Silent Kappa

Recombinant Human IgG1 Monoclonal Antibody with Fc silent mutation. 
Clone: 3H3.
Isotype: Human IgG1 kappa.
Source: The anti-mouse CD137 (TNFRSF9 or 4-1BB) monoclonal antibody (clone: 3H3) was produced in mammalian cells.
Conjugation: Non-conjugated.
Specificity/Sensitivity: The recombinant 3H3 antibody specifically binds to mouse 4-1BB protein.
Applications: Flow cytometry (FC), and Immunohistochemistry - Frozen (IHC-F). 
Form of Antibody: 0.2 uM filtered solution, pH 7.4, containing 0.09% sodium azide.
Purity: >95% by SDS-PAGE under reducing conditions.

Shipping: The 3H3 antibody is shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 
12 months from date of receipt, 2 to 8°C as supplied. Do not freeze it.

References of anti-mouse CD137 (TNFRSF9 or 4-1BB) antibody (3H3)

Preclinical characterization and phase 1 results of ADG106, a non-fucosylated agonistic anti-CD137 antibody for the treatment of advanced solid tumors and non-Hodgkin's lymphoma.
Wang Z, et al. Cell Rep Med. 2024 Feb 20;5(2):101397. doi: 10.1016/j.xcrm.2024.101397. PMID: 38330942
To assess ADG106 in a murine tumor model, syngeneic mice bearing MC38 tumors were treated with ADG106 or 3H3 antibody. Flow cytometry analysis showed increased CD8+ T cell infiltration in tumors treated with ADG106 compared to 3H3. The antibody 3H3 was used as a positive control for CD137 agonism. Peripheral blood samples were stained for CD137 expression using 3H3. Data were acquired on a BD FACSCelesta flow cytometer.
Tags: anti-mouse CD137 3H3; anti-mouse 4-1BB 3H3

TCR-independent CD137 (4-1BB) signaling promotes CD8+ T cell expansion and effector function.
Liu S, et al. Immunity. 2023 Jul 11;56(7):1465-1482.e6. doi: 10.1016/j.immuni.2023.06.009. PMID: 37392737
Cells were stained with anti-CD137 antibody (clone 3H3, BioLegend) for flow cytometry to assess expression levels. The 3H3 antibody was used to gate CD137+ CD8+ T cells in tumor-infiltrating lymphocytes. Flow cytometry was performed on a BD LSRFortessa. CD8+ T cell expansion was measured by CFSE dilution in the presence of 3H3 stimulation. Data analysis was conducted using FlowJo v10.
Tags: function of 3H3 antibody; 3H3 antibody

Soluble CD137 as a dynamic biomarker to monitor agonist CD137 immunotherapies.
Gavilán A, et al. J Immunother Cancer. 2022 Mar;10(3):e003532. doi: 10.1136/jitc-2021-003532. PMID: 35236742
CD137 agonist therapy was provided by 5 μg of anti- CD137 (clone 3H3, BioXcell) intratumorally administered three times with a 3- day interval. Similarly, isolated mouse CD8 T cells from splenocytes were activated for 72 hours in plates coated with the mAbs anti- CD3 ε (17A2, 0.5 μg/mL), anti- CD137 (3H3, 10 μg/ mL), anti- CD28 (37.51, 5 μg/mL) and/or RatIgG2a isotype control (RTK2758). Agonist anti- CD3 ε (OKT3 and 17A2), anti- CD137 (6B4 and 3H3), mIgG1 isotype control (MOPC- 21), RatIgG2a isotype control (RTK2758) mAbs and human Mesothelin- Fc protein (ACROBiosystems) were covalently coupled to Dynabeads M- 450 Tosylactivated (ThermoFisher) according to the manufacturer’s instructions. OKT3 tagged with AlexaFluor647 and 6B4 tagged with AlexaFluor488 were used to visualize by Flow Cytom- etry the graded density of anti- CD137 antibody coupled to the beads. Flow cytometry was used to monitor CD137 surface expression.
Tags: 3H3 antibody for flow cytometry; 3H3 mAb for cancer research

Differentiated agonistic antibody targeting CD137 eradicates large tumors without hepatotoxicity.
Madireddi S, et al. JCI Insight. 2020 Mar 12;5(5):e133647. doi: 10.1172/jci.insight.133647. PMID: 32161196
In order to understand the effect of epitope and in vitro potency on in vivo activity, we compared CTX-471-AF with the urelumab-like mouse surrogate antibody 3H3. To assess the potential of CTX-471, CTX-471-AF, and 3H3 to induce liver inflammation, we administered up to 80 mg/kg of these antibodies to nontumor-bearing mice by weekly i.v. injections. Comparison studies were performed against anti-CD137 clone 3H3 and a panel of well-characterized immuno-oncology antibodies targeting PD-1, PD-L1, CTLA-4, or OX40 that have demonstrated activity against smaller CT-26 tumors in the literature (Figure 8, B and C). The 3H3 antibody displayed superagonist activity in the mouse assay, further supporting its use as a functional surrogate for urelumab. In the CT-26 murine colon carcinoma model, both CTX-471-AF and CTX-471 demonstrated potent antitumor activity across a broad dose-range in mice bearing established (50–75 mm³) tumors (Figure 4A and Supplemental Figure 3A), resulting in a high rate of complete cures and significantly improved overall survival (Figure 4B and Supplemental Figure 3B).
Tags: 3H3 mAb for mouse tumor model; 3H3 monoclonal antibody

CD137/OX40 Bispecific Antibody Induces Potent Antitumor Activity that Is Dependent on CD8+ T Cells.
Gasparian S, et al. Cancer Immunol Res. 2021 Mar;9(3):325-338. doi: 10.1158/2326-6066.CIR-20-0580. PMID: 32273279
A different anti-mouse CD137 (clone 3H3) was also tested in this assay and showed crosslink-independent activity, similar to that seen with Fab clone 20H4.9 (Fig. 3A and Supplemental Fig. S6A and S6B). The results showed a clear difference between the two CD137 agonist antibodies tested, clone 3H3 induced a sustained increase in T cell infiltration, proliferation and activation in the liver and spleen, whereas clone Lob12.3 did not (Fig. 6B). For cell binding assays cells were incubated with primary antibodies or mAb 2 followed by detection with an anti-human Fc-488 secondary antibody (Jackson Immunoresearch). Cells isolated from tissues and blood were stained for flow cytometry with fluorochrome conjugated antibodies including those against CD4, Ki67, FoxP3, CD69, CD3, CD8 (Thermo), CD45 (BD) and fixable live/dead dye (Thermo) in the presence of Fc block (Thermo) according to manufacturer’s recommendations. TIL analysis of day 21 CT26 tumors (n=5) treated with 1 mg/kg FS120 surrogate or controls Q2D starting on day 10 for 3 doses injected intraperitoneally by flow cytometry.
Tags: anti-mouse CD137 3H3 antibody; anti-mouse CD137 3H3 for flow cytometry

For more references about anti-mouse CD137 (TNFRSF9 or 4-1BB) antibody (3H3), please contact our scientific support team with message@sydlabs.com.

Syd Labs provides the following in vivo grade recombinant anti-mouse CD137 (TNFRSF9 or 4-1BB) monoclonal antibodies:

Recombinant Anti-mouse CD137 (TNFRSF9 or 4-1BB) monoclonal antibody (Clone: 3H3)