Mouse procedures and studies were approved by the Vanderbilt Division of Animal Care and the Institutional Animal Care and Use Committee. Established MMTV-Neu mammary tumor cells or primary MMTV-polyoma V middle-T mammary tumor cells (1 × 106) were orthotopically injected into the 4th mammary fat pads of FVB/n mice (or athymic nu/nu mice, for MMTV-Neu). Following the establishment of tumors (~100–200 mm3), the mice were treated with diluent or guadecitabine (3 mg/kg, I.P. injection, 3 continuous days), or in combination with isotype IgG (BioXcell, clone LTF-2, 100 µg intraperitoneal, on days 3, 7, 10, 14) or α-PD-L1 (BioXcell, clone 10F. 9G2, 100 µg intraperitoneal, on days 3, 7, 10, 14). For T cell infiltration analysis, mice were euthanized on Day 7 after the initiation of treatment and tumor samples were collected for IHC. Six to eight (6–8) mice were used for each treatment arm for these studies. For tumor growth analysis, tumor was measured 2–3 times weekly with calipers and volume was calculated in mm3 using the formula (length x width x width/2). Mice were humanely euthanized when the tumor volume reached 2 cm3 or 5 weeks initiation of treatment (4–5 weeks for the combination study). At least five mice were used for each treatment arm for tumor growth studies.
Ltf 2
Manufactured by BioXCell
Sourced in United States
The LTF-2 is a laboratory centrifuge designed for general-purpose applications. It can be used to separate and isolate a variety of biological samples, including cells, organelles, and macromolecules. The LTF-2 features a compact and durable design, allowing for reliable and efficient operation in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
Rb6 8c5, by BioXCell (6 mentions)
Rmp1 14, by BioXCell (5 mentions)
Yts 169, by BioXCell (4 mentions)
Clone gk1, by BioXCell (3 mentions)
Anti cd4 gk1, by BioXCell (2 mentions)
Anti cd8 2.43, by BioXCell (2 mentions)
Rat igg2b, by BioXCell (2 mentions)
Cd4 pe cy7, by BioLegend (2 mentions)
Fixable aqua dead cell stain, by Thermo Fisher Scientific (2 mentions)
Cd3 alexa 488, by BioLegend (2 mentions)
Fix lyse solution, by Thermo Fisher Scientific (2 mentions)
Cd8b precp cy5, by BioLegend (2 mentions)
Ack lysis buffer, by Thermo Fisher Scientific (2 mentions)
Cd45 buv395, by BD (2 mentions)
Mar1 5a3, by BioXCell (2 mentions)
Anti gr1 clone rb6 8c5, by BioXCell (2 mentions)
Diphtheria toxin, by Merck Group (2 mentions)
Clone cxcr3 173, by BioXCell (1 mentions)
Clone 2a3, by BioXCell (1 mentions)
C57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
77 protocols using ltf 2
1
Combination Therapy for Mammary Tumor Regression
2
Neutrophil Depletion in Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-GR1 (Bio X Cell, clone RB6-8C5) or a rat IgG2b isotype control (Bio X Cell, clone LTF2) antibodies were injected intraperitoneally from 1 to 3 days post-infection (0.25 mg per mouse and per day), in saline. The efficacy of the depletion was assessed by histology and flow cytometry (data not shown).
3
T Cell Depletion and Recruitment Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD8+ and CD4+ T cell depletions were accomplished with mouse anti-CD8 (BioXcell, Clone 2.43) and mouse anti-CD4 (BioXcell, Clone GK1.5), respectively. Depleting antibodies were given via i.p. injection at 200 μg/injection every two days starting on D6. For T cell recruitment studies, mouse anti-CXCR3 antibody (BioXcell, Clone CXCR3-173) was administered i.p, at 200 μg/injection. Prevention of T cell LN egress was performed with fingolimod hydrochloride (FTY720; Cayman, 10006292) administered i.p. every other day at 2 mg/kg. All antibody experiments utilized isotype controls (BioXcell, Clone 2A3 or LTF-2).
4
T-cell Adoptive Transfer for Immunotherapy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C57BL/6 mice and BALB/c nude mice were purchased from Japan Clea (Tokyo, Japan). Sult2b1–/– mice were obtained from the Jackson Laboratory (stock no. 018773, Bar Harbor, ME, USA), which had been backcrossed with C57BL/6 mice more than seven generations before use (21 (link)). DOCK2–/– mice on a C57BL/6 genetic background have been described previously (13 (link)) and were used in some experiments after crossing with OTII T-cell receptor (TCR) Tg mice. Mice were maintained under specific pathogen-free conditions in the animal facility of Kyushu University. The animal experiment protocol was approved by the committee of Ethics of Animal Experiments, Kyushu University. When indicated, 200 µl of anti-mouse PD-L1 (programmed cell death ligand 1) antibody (10F.9G2, 1 mg ml−1, BioXCell, West Lebanon, NH, USA) or isotype-matched control (LTF-2, 1 mg ml−1, BioXCell) was injected intra-peritoneally into tumor-bearing mice. For T-cell transfer, activated OTII CD4+ T (2 × 106) and OTI CD8+ T cells (5 × 105) were injected intravenously into tumor-bearing mice.
5
Orthotopic HCC Model for Immunotherapy
6
Nephrotoxic Serum-Induced Kidney Injury
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NTN was induced in male mice (8–12 weeks old) by intraperitoneal (i.p.) injection of 2.5 mg nephrotoxic sheep serum per gram of mouse body weight as described previously46 (link). Mice were sacrificed 8 days after NTN induction. To assess systemic antibody response, heart blood was drawn from individual mice. One day before killing, mice were housed in metabolic cages for urine collection. Albuminuria was determined by ELISA (Mice-Albumin Kit; Bethyl, Montgomery, TX). C57BL/6 mice were treated i.p. with an anti-PD-L1 antibody (250 µg/mouse; 10 F.9G2; BioXCell, West Lebanon, NH) daily starting one day after NTN induction. As isotype control, mice received rat IgG2b (250 µg/mouse; LTF-2; BioXCell). PD-L1−/− mice received i.p. an anti-IFNγ antibody (250 µg/mouse; XMG1.2; BioXCell) one day before and 4 days after NTN induction. As isotype control, rat IgG1 (1 mg/mouse; HRPN; BioXCell) was injected.
7
Evaluating APR-246 and Immunotherapies in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
APR-246 was provided by Aprea Therapeutics under a material transfer agreement. APR-246 was reconstituted in PBS just prior to injection and used at 100 mg/kg per mouse administered intraperitoneally (i.p.) daily as depicted in respective experiments; PBS was used as vehicle control. Therapeutic in vivo monoclonal antibodies (mAbs) anti–PD-1 (RMP1-14) and anti–CTLA-4 (9D9), corresponding IgG isotype controls (2A3 and MPC-11), and depleting mAbs anti-CD4 (GK1.5) and anti-CD8 (2.43) and their IgG isotype controls (LTF-2) were purchased from Bio X Cell. RMP1-14 (250 μg) and 2A3 (250 μg) were administered i.p. twice weekly beginning on day 7 for up to 4 doses. Depleting mAbs GK1.5 (560 μg) and 2.43 (400 μg) were administered i.p. twice weekly beginning on day 7 for 4 doses.
8
Anti-CD4 Antibody Treatment in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
9
In vivo T-cell depletion and serum transfer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4 or CD8 T-cell subsets were depleted in BALB/c mice by intraperitoneal injection of 250 µg of anti-CD4 (rat IgG2b, Clone GK1.5, Bio X Cell) or anti-CD8 (rat IgG2b, Clone 53-6.7, Bio X Cell) antibody 1 day before and 1 day after challenge. As control, an isotype-matched antibody (rat IgG2b, LTF-2, Bio X Cell) was used. For long-term depletion of CD4 T cells, injection was performed 2 days before prime vaccination and twice a week thereafter. At Day 39 (4 days post RSV-A2 challenge), the efficacy of these treatments was determined by FACS analysis of heparinized whole blood and splenocytes. Briefly, cells were treated with a purified anti-mouse CD16/CD32 monoclonal antibody (1:50, Fc Block™, BD) prior to staining with CD8 FITC (1:200; BioLegend) and CD4 brilliant violet 785 (1:200, BioLegend). For live versus dead cell discrimination, Zombie Aqua™ (1:200, BioLegend) was used. All cells were acquired using a digital flow cytometer (LSR II, BD Biosciences), and data were analyzed with FlowJo software (Tree Star).
Serum from MVA-RSV-vaccinated or mock-treated control mice was transferred by intraperitoneal (IP) injection of 1 ml of serum to non-vaccinated BALB/c mice 1 day before challenge.
Serum from MVA-RSV-vaccinated or mock-treated control mice was transferred by intraperitoneal (IP) injection of 1 ml of serum to non-vaccinated BALB/c mice 1 day before challenge.
10
Antibody characterization and in vivo use
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 314.8 mAb (mouse IgG1 anti-human ICOS) has been described before.21 Isotype controls (mouse IgG1, MOPC-1; rat IgG2b, LTF-2) and anti-Gr1 mAb (rat IgG2b, RB6-8C5) were purchased from BioXcell (West Lebanon, NH, USA). The MT807R1 recombinant Ig consisting of rhesus IgG1k constant regions and CDRs derived from the anti-human CD8 antibody M-T807 grafted into rhesus variable framework regions and was provided by the Nonhuman Primate Reagent Resource (NIH contract HHSN272200900037C and grant RR016001). The antibody was expressed in vitro using serum-free medium and purified by protein-A affinity chromatography. Endotoxin was <1EU/mg. Cyclophosphamide (CTX, Sigma Aldrich) was prepared extemporaneously according to supplier technical data sheet, i.e to 20 mg/ml of injectable water. All reagents were injected intraperitoneally.
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!