BALB/c or C57BL/6 FoxP3DTR mice were injected in the mammary fatpad or subcutaneously (s.c.) with either 4T1.2 (1 x 105) or MC38 cells (1×106), respectively. Tumor growth was measured regularly by caliper square measurements and treatment started when the mean tumor size was over 40 mm2. Mice were euthanized when tumor size reached 150 mm2. To deplete Tregs and uncover antibody treatment toxicity, mice were injected intraperitoneally with 250 ng of diphtheria toxin (DT) (Sigma-Aldrich, St Louis, USA), diluted in PBS. Some mice additionally received treatment at the indicated dose and schedule with 200 µg rat control IgG2a (1-1; Leinco Technologies, St. Louis, USA), 100 µg anti-CD40 (FGK4.5, BioXCell, West Lebanon, USA), 200 µg anti-TNFα (TN3-19.12; Leinco Technologies) and 500 µg anti-interleukin (IL)-6R (MR16-1; Chugai Pharma, Tokyo, Japan).
Anti cd40 fgk4
Manufactured by BioXCell
Sourced in United States
Anti-CD40 (FGK4.5) is a monoclonal antibody that binds to the CD40 receptor on the surface of B cells and other antigen-presenting cells. It is a laboratory tool used in immunological research applications.
Automatically generated - may contain errors
Lab products found in correlation
Poly 1 c, by InvivoGen (3 mentions)
Poly 1 c, by Cytiva (2 mentions)
Pierce limulus amebocyte lysate chromogenic endotoxin quantitation kit, by Thermo Fisher Scientific (2 mentions)
Pam3csk4, by InvivoGen (2 mentions)
Lipopolysaccharides (lps), by InvivoGen (2 mentions)
Diphtheria toxin, by Merck Group (1 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions)
Tnf α, by BioLegend (1 mentions)
Anti cd40, by Merck Group (1 mentions)
Recombinant mouse il 4, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by Merck Group (1 mentions)
Polybrene, by Merck Group (1 mentions)
Mitomycin, by Merck Group (1 mentions)
Anti cd28 37, by BioXCell (1 mentions)
Mouse cd8 t cell isolation kit, by STEMCELL (1 mentions)
Murine baff, by R&D Systems (1 mentions)
Anti biotin microbead, by Miltenyi Biotec (1 mentions)
Mpc 11, by BioXCell (1 mentions)
Anti ctla4 9h10, by BioXCell (1 mentions)
Alf 161, by BioXCell (1 mentions)
15 protocols using anti cd40 fgk4
1
Immune Modulation in Murine Tumor Models
2
Peptide vaccination with polyI:C and anti-CD40
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eight weeks HHD-DR1 mice were vaccinated with individual peptides (100 nmoles/mouse) combined with polyI:C (Amersham) and anti-CD40 (FGK4.5; Bioxcell), both being administered at 50 µg/mouse. Class I-restricted peptides were administered subcutaneously, whereas class II-restricted peptides were administered intravenously and were also given in a boost 1 week later to increase their immunogenicity. Seven days after the last immunization, mice were sacrificed, and spleens obtained.
3
Melanoma and Triple-Negative Breast Cancer Mouse Models
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For melanoma tumor studies, 2.5×105 B16F10 cells in 50 μL DMEM were injected s.c. into C57BL/6 mice above the right flank. For a triple-negative breast cancer model, 2×105 4T1 cells in 30 μL DMEM were injected into mammary fat pad number five. Tumor size was measured twice a week using calipers. For immunotherapy, tumor-bearing mice were injected intratumorally twice, 2 days apart, with 100 μg anti-CD40 (FGK4.5; BioXCell), 0.2 μg TNFα (BioLegend), and 200 μg/mouse anti-TRP1 antibody (TA99; BioXCell). For adoptive cell transfer, splenic T cells were infected with pMIGII encoding TCR recognizing MHCI-gp10025-33(von Boehmer et al., 2013 (link)), or MHCI-TRP2180-188(Tran et al., 2016 (link)), or MHCII-TRP1113-126(Yost et al., 2019 (link)). Recipient mice were sub-lethally irradiated at a single dose of 600 rad, and injected i.v. twice, 3 days apart, with 1 × 106 transduced T cells followed by i.p. injections of 300,000 IU of IL2 (PeproTech) for 4 consecutive days.
4
Retroviral Transduction of BAFFR in SW HEL B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
cDNA encoding membrane-bound BAFFR protein was subcloned between the NotI and XhoI sites of the plasmid murine stem cell virus (IRES)–internal ribosome entry site–GFP vector (Gatto et al., 2009 (link)), and retroviral particles from empty and BAFFR-expressing vectors were generated using the Phoenix-E packaging cell line together with the pCL-Eco packaging vector as described previously (Swift et al., 2001 ). For retroviral transduction of SWHEL B cells, spleen cells were harvested and stimulated in vitro with recombinant mouse IL-4 (10 ng/ml; R&D Systems) and anti-CD40 (FGK4.5, 1 µg/ml; BioXcell) in RPMI supplemented with 10% FCS, 55 mM mercaptoethanol, 1 mM sodium pyruvate, 0.1 M nonessential amino acids, 10 mM Hepes, 50 U/ml penicillin, 50 mg/ml streptomycin) for 20 h. Spin transduction of stimulated B cells was then performed with retroviral supernatants in the presence 4 µg/ml polybrene (Sigma-Aldrich) for 4 h and restimulated with medium containing IL-4 and anti-CD40 for a further 20 h. Cells were washed with medium, and the efficiency and effectiveness of transduction were analyzed by flow cytometric analysis of GFP expression and cell surface BAFFR expression.
5
Vaccinia and ova-psDNA Immunization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
6–8-week-old C57BL/6 (CD45.2) mice were immunized with 1E3 or 1E4 colony-forming units (CFU) of Vaccinia Western Reserve or 5 µg of poly I:C (Invivogen) with or without 5 µg of anti-CD40 (FGK4.5, BioXcell)and 10 µg of ova-psDNA or ova in 50 μL volume by footpad injection. Endotoxin levels were quantified using the Pierce Limulus Amebocyte Lysate Chromogenic Endotoxin Quantitation kit (Thermo Scientific) to be less than 0.5 EU/mg for either ova or ova conjugated to psDNA.
6
T Cell Activation Assay with DCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Purified DCs (105) were stimulated with one of either PolyIC, CpG, LPS or Pam3Csk4 (all from Invivogen), or anti-CD40 (FGK4.5, BioXcell). CD4+ T cells (105) were stimulated with α-CD3 (2C11 hybridoma supernatant), in the presence of mitomycin-c-treated (50µg/ml, Sigma) DCs (105) or stimulated with α-CD3 and α-CD28 (37.51, BioXcell) without DCs for 2 days. For DC co-culture with BDC2.5 CD4+ T cells, mitomycin-c-treated DCs (105) were co-cultured with BDC2.5 CD4+ T cells in a 1:1 ratio in the presence of BDC2.5 mimotope peptide (RTRPLWVRME) (39 (link)). After a 48-hour incubation, the culture supernatants were collected followed by pulsing the cells with 3H-thymidine. Cells were incubated for an additional 18 hours before harvesting. Proliferation was determined by 3H-Thymidine incorporation using a β-counter and are presented as counts per minute (CPM), after subtracting background (cells without stimulation), termed ΔCPM.
7
Lymphocyte Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total spleen cells or purified B cells were stimulated with anti-CD40 (FGK4.5, BioXcell), with anti-IgM, or anti-CD3 (2C11 BioXcell) and anti-CD28 (37.51, BioXcell), or CpG (Invivogen), LPS (Invivogen) and Pam3Csk4 (Invivogen) at 37 °C, 5% CO2 for 3 days. The proliferative response was detected by 3H-thymidine incorporation and read on a β-counter. The cell proliferation was presented as counts per minute (cpm).
8
Generation of Tumor-Induced B Vax Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BVax were generated from 4-1BBL+ B cells from spleens and dCLNs of tumor-bearing mice. Mice were challenged with 2 × 106 tumor cells and were sacrificed 12–14 d after tumor inoculation. B cells were negatively isolated from spleens and dCLNs using the EasySep Mouse B Cell Isolation Kit. 4-1BBL+ cells were then magnetically positively isolated using anti-mouse 4-1BBL biotin (5F4; BioLegend) and antibiotin microbeads (Miltenyi Biotec). Cells were resuspended at 2 × 106 cells/ml of complete RPMI, supplemented with 100 nM of murine BAFF (R&D), and activated with anti-CD40 (FGK4.5; BioXCell). 24 h later, 10 U/ml of murine IFNγ was added to the culture. 48 h after the time of isolation, BVax were harvested, counted, and ready for further utilization. In many experiments, BVax were concomitantly injected with CD8+ T cells that also originated from spleens and CLNs of tumor-bearing mice, isolated using the Mouse CD8+ T Cell Isolation Kit (StemCell Technologies).
9
In Vivo Immunotherapy Combination Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
APR-246 (2 mg) was dissolved in PBS and administered at 100 mg/kg intraperitoneally. Therapeutic in vivo mAbs anti-CD40 (FGK45) and corresponding IgG isotype control (2A3), anti-CTLA-4 (clone 9D9) or the matched IgG isotype control (MPC11) were purchased from Bio X Cell. 9D9 (100 μg) and MPC11 (100 μg) were administered intraperitoneally twice weekly. FGK45 (20 μg), MPLA (5 μg), and 2A3 (20 μg) were administered concurrently intratumorally twice weekly. MPLA (Sigma-Aldrich) was reconstituted as previously described (Stark et al, 2015 (link)). On day 0 of the experiments, tumor cells were implanted intradermally (i.d) in the flank (right if a unilateral model was used). 2.5 × 105 B16 Trp53 KO or 5 × 105 B16-WT were injected into C57BL/6J mice. PBS was used to resuspend the cells at the appropriate dilution. Treatments were started at D7 post tumor implantation, as single agents or in combinations for 3 wk with the appropriate regimen for each drug. Tumors were measured twice a week with a caliper, and the tumor size was calculated based on an ellipsoid formula. Mice that had no visible and palpable tumors on consecutive measurement days were considered complete regressions. Animals were euthanized for signs of distress or when the total tumor volume reached 2,500 mm3.
10
Vaccinia and CHIKV Infection Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
6–8-week-old C57BL/6 (CD45.2) mice were immunized with 1E4 plaque-forming units (PFU) of Vaccinia Western Reserve or 5 μg of poly I:C (Invivogen) with or without 5 μg of anti-CD40 (FGK4.5, BioXcell) and 20 μg of ova-psDNA or ova in 50 μL volume per footpad injection. Endotoxin levels were quantified using the Pierce Limulus Amebocyte Lysate Chromogenic Endotoxin Quantitation kit (Thermo Scientific) to be less than 0.5 EU/mg for either ova or ova conjugated to psDNA. For CHIKV infections, 4 week old C57BL/6 mice were inoculated with 103 PFU of CHIKV 181/25 or CHIKV AF15561 (WT CHIKV) in a 10 μL volume in both rear footpads.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!