Invivomab rat igg2a isotype control

Manufactured by BioXCell

Sourced in United States

InVivoMAb rat IgG2a isotype control is a laboratory reagent used as a control in experimental procedures. It is a rat immunoglobulin G (IgG) of the IgG2a subclass, which can be used to assess non-specific binding or background signal in experiments involving the use of rat-derived antibodies.

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Lab products found in correlation

Invivomab anti mouse pd 1 cd279, by BioXCell (5 mentions) Takinib, by Cayman Chemical (1 mentions) Anti mouse cd8α, by Cell Signaling Technology (1 mentions) 5z 7 oxozeaenol, by Enzo Life Sciences (1 mentions) Flag tag (flag), by Cell Signaling Technology (1 mentions) Gsdmd, by Cell Signaling Technology (1 mentions) Gsdme, by Abcam (1 mentions) Invivomab mouse igg2a isotype control, by BioXCell (1 mentions) Anti nk1, by BioXCell (1 mentions) Invivopure ph 7.0 dilution buffer, by BioXCell (1 mentions) Invivomab rat igg2b isotype control, by BioXCell (1 mentions) Invivomab anti mouse pd l1 b7 h1, by BioXCell (1 mentions) Bibf 1120, by Boehringer Ingelheim (1 mentions) Anti cd11b apc, by BD (1 mentions) Anti mouse cd16 cd32, by BD (1 mentions) Invivomab anti mouse ly6g, by BioXCell (1 mentions) Anti rabbit igg hrp, by Beyotime (1 mentions) Cy3 donkey anti mouse igg h l, by Jackson ImmunoResearch (1 mentions) Anti mouse f4 80 pe, by BD (1 mentions) Anti mouse ly6g pe, by BD (1 mentions)

10 protocols using invivomab rat igg2a isotype control

Establishment and Evaluation of Murine Tumor Models

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Cells (5.0 × 10⁵ cells in 100 μL phosphate-buffered saline) were mixed with 100 μL Matrigel Matrix Basement Membrane High Concentration (Corning, Inc., Corning, NY). The cells were injected subcutaneously into 8-week-old male NOD.CB17-Prkdcscid/J or C57BL/6 mice. For the establishment of orthotopic liver tumor models, the cells (1.0 × 10⁶ cells in 100 μL phosphate-buffered saline) were injected via the spleen into 8-week-old male C57BL/6 mice. At 30 days after injection, hepatic tumors were evaluated by hematoxylin and eosin staining and immunohistochemistry.
Purified anti-mouse PD1 monoclonal antibody, InVivoMAb anti-mouse PD-1 (CD279) (catalog no. BE0146, clone: RMP1-14), and control Ig, InVivoMAb rat IgG2a isotype control (catalog no. BE0089, clone: 2A3), were purchased from BioXCell (Lebanon, NH). The anti-PD1 antibody (100 μg/mouse) was intraperitoneally administered to MHCF1- or MHCF5-derived tumor-bearing mice every 3 days from day 6 to 18 (total 5 times), and tumor volumes were evaluated until 60 days and 25 days, respectively.

Shirasaki T., Murai K., Honda M., Okada H., Innami Y., Yamada A., Shimakami T., Kawaguchi K., Yamashita T., Sakai Y, & Kaneko S. (2021). Establishment of liver tumor cell lines from atherogenic and high fat diet fed hepatitis C virus transgenic mice. Scientific Reports, 11, 13021.

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Melanoma Xenograft Immunotherapy in Mice

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For the in vivo treatment model, 1.5 × 10⁶ YUMMER1.7 cells in 50 µL of PBS were subcutaneously inoculated in the right hind paw of 6-week-old C57BL-6 mice. Cells were harvested by trypsinization and resuspended in PBS (pH 7.4) before animals were injected.
During inoculation, mice were kept under inhalational anesthesia with isoflurane 2% in 2 L/min airflow.
Tumor-bearing mice were injected intraperitoneally (i.p) with PD1 antibody or isotype control antibodies (10 mg/kg) 3 times per week (days0-2-4-7-9-11) for 2 weeks at a size of 300 ± 50 mm³. Antibodies (in vivo MAb rat IgG2a isotype control and in vivo MAb anti-mouse PD-1 (CD279)) were purchased from BioXcell (Lebanon, NH, USA). After the treatments were interrupted, tumor regrowth was longitudinally monitored using an electronic caliper and calculated via the formula: 4/3 × π×(L/2) × (l/2)². The growth delay of melanoma xenografts was calculated as the time taken (in days) to reach a volume of 1000 mm³.

Farah C., Neveu M.A., Bouzin C., Knezevic Z., Gallez B., Leucci E., Baurain J.F., Mignion L, & Jordan B.F. (2023). Hyperpolarized 13C-Pyruvate to Assess Response to Anti-PD1 Immune Checkpoint Inhibition in YUMMER 1.7 Melanoma Xenografts. International Journal of Molecular Sciences, 24(3), 2499.

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Investigating TAK1 Signaling Pathway

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The commercial antibodies used were ESRP2, β‐actin, TAK1, phospho‐TAK1 (Ser439), GSDME (Abcam), p38, phospho‐p38, JNK, phospho‐JNK, p65, phospho‐p65, IKK, phospho‐IKK, cleaved caspase‐3, TAB1, TAB2, GSDMD, anti‐mouse CD8α, FLAG (Cell Signaling Technology), TAB3 (Santa Cruz Biotechnology), InVivoMAb anti‐mouse PD‐1 (CD279), and InVivoMAb rat IgG2a isotype control (Bio X Cell). The TAK1 inhibitor, takinib, was purchased from Cayman Chemical, and 5Z‐7‐oxozeaenol was obtained from Enzo Life Sciences.

Yan Q., Fang X., Liu X., Guo S., Chen S., Luo M., Lan P, & Guan X. (2023). Loss of ESRP2 Activates TAK1‐MAPK Signaling through the Fetal RNA‐Splicing Program to Promote Hepatocellular Carcinoma Progression. Advanced Science, 11(1), 2305653.

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Neutrophil Depletion in Mice

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WT C57Bl6/J and IL-36γ^−/− mice were injected i.p. with InVivoMAb anti-mouse Ly6G (α-Ly6G) antibody or InVivoMAb rat IgG2a isotype control (Bio X Cell, West Lebanon, NH) to deplete neutrophils using 200 µg of antibody in 200 µl at Day −1 prior to infection and then every other day through the duration of the challenge study. CVLs were analyzed by flow cytometry to confirm the efficiency of neutrophil depletion.

Gardner J.K., Swaims-Kohlmeier A, & Herbst-Kralovetz M.M. (2019). IL-36γ is a key regulator of neutrophil infiltration in the vaginal microenvironment and limits neuroinvasion in genital HSV-2 infection. Journal of immunology (Baltimore, Md. : 1950), 203(10), 2655-2664.

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Selective Immune Cell Depletion for Cancer

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Depletion of NK cells and CD8+ T cells was achieved by administering anti-NK1.1 (PK136, 200 μg) or anti-CD8α (53–6.7, 400 μg), or InVivoMab mouse IgG2a isotype control (BioXCell, C1.18.4, 200 μg), or InVivoMab rat IgG2a isotype control (BioXCell, 2A3, 400 μg) in PBS via intraperitoneal injection every 3 days beginning on day 6 post tumor engraftment. Depletion of Tregs was achieved by administering anti-OX40 (OX86, 200 μg), or PBS via intraperitoneal injection on day 7 and 11 of tumor growth. anti-NK1.1, anti-CD8α, and anti-OX40 antibodies were provided by AstraZeneca.

Dean I., Lee C.Y., Tuong Z.K., Li Z., Tibbitt C.A., Willis C., Gaspal F., Kennedy B.C., Matei-Rascu V., Fiancette R., Nordenvall C., Lindforss U., Baker S.M., Stockmann C., Sexl V., Hammond S.A., Dovedi S.J., Mjösberg J., Hepworth M.R., Carlesso G., Clatworthy M.R, & Withers D.R. (2024). Rapid functional impairment of natural killer cells following tumor entry limits anti-tumor immunity. Nature Communications, 15, 683.

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Murine Immune Checkpoint Blockade Protocol

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The InVivoMab anti-mouse CD8alpha (BE0061), InVivoMab anti-mouse NK1.1 (BE0036), InVivoMab anti-mouse PD-1 (CD279) (BE0273), InVivoMab anti-mouse PD-L1 (B7-H1) (BE0101), InVivoMab rat IgG2a isotype control (BE0089), and InVivoMab rat IgG2b isotype control (BE0090) were purchased from Bio X Cell, diluted in InVivoPure pH 7.0 Dilution Buffer (IP0070), and administered as indicated in the corresponding figures. Rabbit anti-murine RANTES (CCL5) (500-P118) was purchased from PeproTech.

Noman M.Z., Parpal S., Van Moer K., Xiao M., Yu Y., Viklund J., De Milito A., Hasmim M., Andersson M., Amaravadi R.K., Martinsson J., Berchem G, & Janji B. (2020). Inhibition of Vps34 reprograms cold into hot inflamed tumors and improves anti–PD-1/PD-L1 immunotherapy. Science Advances, 6(18), eaax7881.

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Anti-PD-1 Immunotherapy in Mice

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When tumors reached measurable size (ca. 30 mm³), mice were treated with 10 mg/kg of anti-PD-1 (InVivoMAb anti-mouse PD-1 (CD279), Bio X Cell) by intraperitoneal injection. Control animals were treated with 10 mg/kg of IgG2a isotype (InVivoMAb rat IgG2a isotype control, Bio X Cell). After the first injection, mice received injections every third day for a total of approximately 2–4 weeks, depending on the duration of the study.

Frias A., Di Leo L., Antoranz A., Nazerai L., Carretta M., Bodemeyer V., Pagliuca C., Dahl C., Claps G., Mandelli G.E., Andhari M.D., Pacheco M.P., Sauter T., Robert C., Guldberg P., Madsen D.H., Cecconi F., Bosisio F.M, & De Zio D. (2023). Ambra1 modulates the tumor immune microenvironment and response to PD-1 blockade in melanoma. Journal for Immunotherapy of Cancer, 11(3), e006389.

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Anti-TIM-3 Antibody Treatment in Mice

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Mice were fed and infected as described. Additionally from the day of infection on, mice were intraperitoneally injected with 100 µg inVivo MAb anti-mouse TIM-3 antibody (BioXCell; BE0115) or InVivoMAb rat IgG2a isotype control (BioXCell; BE0089) (30 (link)–34 (link)) in 200 µl PBS every second day till day 14 post infection.

Pfeifhofer-Obermair C., Tymoszuk P., Nairz M., Schroll A., Klais G., Demetz E., Engl S., Brigo N, & Weiss G. (2021). Regulation of Th1 T Cell Differentiation by Iron via Upregulation of T Cell Immunoglobulin and Mucin Containing Protein-3 (TIM-3). Frontiers in Immunology, 12, 637809.

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Evaluating Antiangiogenic Compounds in Mice

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To evaluate different antiangiogenic compounds, mice were randomized to four separate intervention groups with 12 mice per group (control IgG group n = 14). Treatment started one day prior to heart injection to ensure full BM preventive activity and was always adapted to body weight.
The first group received daily treatment with Nintedanib (BIBF 1120, Boehringer-Ingelheim) in comparison to its control group, receiving 200µL of carrier solution (0,5%-Hydroxyethylcellulose, cat. no.: 822068, Merck) only. Nintedanib is a triple angiokinase inhibitor blocking VEGFR, PDGFR and FGFR kinase activity and was shown to reduce vessel density and vessel integrity in human tumor xenografts [27 (link)]. It was solved in 0,5%-Hydroxyethylcellulose (final concentration 5 mg/mL) and applied via oral gavage in a dosage of 50 mg/kg (ca. 200 µL per mouse).
The third group was treated every 3rd day with a combined anti-VEGF and anti-Ang2 nanobody (BI836880, MW appr. 40.7 kDa; obtained by Boehringer-Ingelheim) in contrast to its respective control group (fourth group), which received a control antibody (InVivoMAb rat IgG2a isotype control, MW 150 kDa; BioXCell) of equal dosage, frequency and concentration. Nanobody and control antibody were solved in sterile PBS (cat. no: D8537, Sigma Life Sciences) reaching a concentration of 2.615 mg/mL, their application dose was 15 mg/kg (100–150µL per mouse).

Kovalchuk B., Berghoff A.S., Karreman M.A., Frey K., Piechutta M., Fischer M., Grosch J., Heiland S., Breckwoldt M.O., Hilberg F., Wick W, & Winkler F. (2020). Nintedanib and a bi-specific anti-VEGF/Ang2 nanobody selectively prevent brain metastases of lung adenocarcinoma cells. Clinical & Experimental Metastasis, 37(6), 637-648.

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Multiparametric Flow Cytometry Analysis of Murine Immune Cell Populations

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The following flow antibodies were purchase from Biolegend: Antimouse CD45-Brilliant Violet 421; Antimouse Ly-6G-Alexa Fluor 488; Antimouse Ly-6G-Alexa Fluor 647; Antimouse CD31-Alexa Fluor 647; Antimouse CD31-PE/Cyanine7; Antimouse CD45 FITC; Anti-mouse/human CD11b-Brilliant Violet 421; Antimouse CD184 (CXCR4)-PerCP/Cyanine5.5; Antimouse CD182 (CXCR2)-APC/Cyanine7.
The following flow antibodies were purchase from Invitrogen: Anti-CD144 (VE-cadherin)-eFluor 660; Anti-CD54 (ICAM-1)-PE; Anti-CD11b-APC; Antimouse F4/80-PE.
The following flow antibodies were purchase from BD Biosciences: Antimouse Ly6G-PE; Anti-Mouse CD16/CD32. Anti-Fibrin (Cat#MABS2155) was purchased from Sigma-Aldrich; Anti-SOD2/MnSOD (Cat#ab68155) and Anti-CD15 (Cat#ab135377) were was purchased from Abcam; Antirabbit β-Actin-HRP (Cat#AF5006) was purchased from Beyotime; Antirabbit IgG-HRP (Cat#7074 S), anti-Ly6G (Cat#87048 S) and anti-COX IV (Cat#4844 S) were purchased from Cell Signaling Technology; Anti-TOMM22 (Cat#66562-1-Ig) was purchased from Proteintech; InVivoMAb antimouse Ly6G (Cat#BE0075-1) and InVivoMAb rat IgG2a isotype control (Cat#BE0089) were purchase from Bio X Cell; Donkey antirabbit IgG (H + L)-Cy3 (Cat#711-165-152) and donkey anti-mouse IgG (H + L)-Cy3 (Cat#715-165-150) were purchased from Jackson ImmunoResearch.

Bao W., Xing H., Cao S., Long X., Liu H., Ma J., Guo F., Deng Z, & Liu X. (2022). Neutrophils restrain sepsis associated coagulopathy via extracellular vesicles carrying superoxide dismutase 2 in a murine model of lipopolysaccharide induced sepsis. Nature Communications, 13, 4583.

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