Beas-2B, HCC827, PC9, A549, H23, H2030, and Jurkat cell lines were maintained in DMEM or RPMI1640 (HyClone) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin-streptomycin (Gibco). The recombinant mouse IL-10 protein (#417-ml-100CF) and recombinant human IL-10 protein (#217-IL-010) was purchased from R&D Systems (Minneapolis, MN, USA). Human EGF (# 100-15-100) was purchased from Peprotech (Cranbury, NJ, USA). Gefitinib (ZD1839) was purchased from Selleck. Anti-PD-1 mAb (#BE0146, clone: RMP1-14), IgG control (#BE0089, clone: 2A3), InVivoPure pH 7.0 Dilution Buffer (#IP0070) and InVivoPure pH 6.5 Dilution Buffer (#IP0065) were purchase from BioXcell (West Lebanon, New Hampshire, USA).
Invivopure ph 6.5 dilution buffer
Manufactured by BioXCell
Sourced in United States
InVivoPure pH 6.5 Dilution Buffer is a laboratory solution designed to maintain a pH of 6.5. It is intended for use in various in vivo and in vitro applications that require a specific pH environment.
Automatically generated - may contain errors
Lab products found in correlation
Invivopure ph 7.0 dilution buffer, by BioXCell (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Gefitinib zd1839, by Selleck Chemicals (1 mentions)
Anti pd 1 mab, by BioXCell (1 mentions)
Control igg, by BioXCell (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Azd6738 atri, by AstraZeneca (1 mentions)
Ah1 peptide spsyvyhqf, by AnaSpec (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Roswell park memorial institute (rpmi), by Lonza (1 mentions)
Be0090, by BioXCell (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Invivomab rat igg2a isotype control anti trinitrophenol, by BioXCell (1 mentions)
5 protocols using invivopure ph 6.5 dilution buffer
1
Cell Line Maintenance and Recombinant Protein Procurement
2
Investigating Immunomodulation in Murine Cancer Models
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CT26 (CRL-2638) and B16-F10 (CRL-6475) were from ATCC and cultured in RPMI (Lonza or Gibco) or DMEM (Lonza), respectively, containing 10 % FBS (Gemini Bio) and 100 U/mL penicillin plus 100 mg/mL streptomycin (Lonza). Cells were tested for mycoplasma. AZD6738 (ATRi) was provided by AstraZeneca and dosed at 75 mg/kg by oral gavage as described previously (15 (link), 17 (link)). FTY720 (Fingolimod hydrochloride, MilliporeSigma) was dissolved in sterile normal saline (0.9% sodium chloride) and dosed at 1 mg/kg via intraperitoneal injection. InVivoPlus anti–mouse PD-L1 antibody (clone 10F.9G2, Bio X Cell) was diluted to 1 mg/mL in InVivoPure pH 6.5 Dilution Buffer (Bio X Cell) and dosed at 10 mg/kg via intraperitoneal injection. AH1 peptide (SPSYVYHQF, Anaspec) was dissolved in DMSO at 20 mg/mL and diluted to a final concentration of 10 μg/mL in T Cell Media (RPMI, 10 % FBS, 100 U/mL penicillin plus 100 mg/mL streptomycin, and the following [all from Gibco]: 1× MEM–nonessential amino acids, 1 mM sodium pyruvate, 5 mM HEPES pH 8.0, 50 mM β-mercaptoethanol).
3
Anti-PD-L1 Antibody Treatment in H. felis Infection
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Anti-mouse PD-L1 (B7-H1) antibody (BE0101; Bio X Cell, West Lebanon, NH) was administered intraperitoneally to H felis–infected mice to block the PD-1/PD-L1 pathway during H felis infection. A total of 300 μg PD-L1 antibody diluted in InVivoPure pH 6.5 Dilution Buffer (IP0065; Bio X Cell) was administered 6 times, once every 3 days, before necropsy. A rat IgG2b isotype antibody (BE0090; Bio X Cell) diluted in InVivoPure pH 7.0 Dilution Buffer (IP0070; Bio X Cell) was used as a control.
4
Generation and Characterization of Murine KL Cell Lines
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Mouse KL cell line was a gift of Ji et al. Somatic KRAS activation and Lkb1 loss were induced in the lungs of genetically engineered male mice (KrasG12D/+/Lkb1L/L with the C57BL6/J genetic background) by nasal inhalation of Adeno-Cre (2 × 106 PFU). After 12 weeks Adeno-Cre treatment, tumors from mice of defined genotypes were dissected and cut into small pieces and cultured in RPMI-1640 + 10% FBS + 1% penicillin–streptomycin. The medium was changed every other day until cells outgrew and stable KL cell lines formed. Mouse KP cell line was provided by InxMed. A549 and H1299 cell lines were purchased from American Type Culture Collection (ATCC). L929 cell line was kindly provided by Stem Cell Bank, Chinese Academy of Sciences. Cells were maintained in DMEM or RPMI1640 (HyClone) supplemented with 10% fetal bovine serum (FBS) (Gibco) and 1% penicillin–streptomycin (Gibco). The small molecule FAK inhibitor IN10018 was provided by InxMed. Anti-PD-1 mAb (#BE0146, clone: RMP1-14), IgG control (#BE0089, clone: 2A3), InVivoPure™ pH 7.0 Dilution Buffer (#IP0070) and InVivoPure™ pH 6.5 Dilution Buffer (#IP0065) were purchase from BioXcell (West Lebanon, New Hampshire, USA).
5
CD8+ T Cell Depletion and EZM8266 Treatment
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On days −3, −2, and −1, mice received a daily i.p. injection of either 500 μg anti-CD8a antibody (BioXCell, Cat. no. BE0004-1; RRID:AB_1107671) or 500 μg InVivoMAb rat IgG2a isotype control, anti-trinitrophenol (BioXCell, Cat. #BE0079). InVivoPure pH 6.5 dilution buffer (BioXCell, Cat. #IP0065) was used as a vehicle for both antibodies. On day 0, CD8 T cell depletion was validated via flow cytometry (Figure S5A –B ), and mice within each antibody group were randomized into four treatment groups based on IVIS total flux: IgG2a + Vehicle control (n = 9), Anti-CD8a antibody (n = 10), 150 mg/kg EZM8266 (n = 9), and Anti-CD8a + EZM8266 (n = 10). EZM8266 was administered daily via oral gavage, and 250 μg anti-CD8a antibody was administered via i.p. injection twice per week starting on day 1.
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