To validate the role of NK cells on the anti-metastatic effect induced by Salmonella YB1, NK cells were depleted in vivo by i.p. injection with 30 µL anti-Asialo-GM1 (#146002; clone Poly21460; BioLegend) antibody on day −1, 1, and 3. NK depletion was maintained by i.p. injection of 20 µL anti-Asialo-GM1 antibody on days 7, 10, and 13. During the experiments, YB1 was injected on day 0 and cancer cells were i.v. injected on day 4. Matched isotype rabbit polyclonal IgG (#BE0095; polyclone; BioXcell) served as the control. NK depletion was confirmed by the absence of CD3− NKp46+ cells in the peripheral blood.
Anti asialo gm1
Manufactured by BioLegend
Anti-Asialo-GM1 is a laboratory reagent used in research applications. It is an antibody that specifically binds to asialo-GM1, a glycolipid present on the surface of certain cell types. This product can be used to identify or isolate cells expressing asialo-GM1.
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Lab products found in correlation
4 protocols using anti asialo gm1
1
Depletion of NK Cells and Antimetastatic Effects
2
Solid and Disseminated Tumor Models in Mice
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For the solid tumor models, 5×106 Raji cells in a 1:1 mixture of endotoxin-free PBS and Matrigel (Corning) were injected s.c. into the flank of CB-17 SCID recipient mice. Mice were randomized when tumor volume reached 50 -100 mm3. Treatment was initiated on day 9.
Alternatively, 5×104 huCD20-B16F10 cells in a 1:1 mixture of endotoxin-free PBS and Matrigel (Corning) were injected s.c. into the flank of μMt- recipient mice (C57BL/6 background). Treatment was initiated the day after implantation and repeated on days 9 and 16.
Tumor size was monitored with a digital caliper (Mitutoyo) every three to four days and is expressed as a volume ((length× width2) /2).
For NK cell depletion, 100 μL of polyclonal anti-asialo-GM1 (Biolegend, Poly21460) antibody was injected i.p. into recipient mice two days before treatment and then once a week. Normal rabbit serum was administered as a control.
For the disseminated tumor model, 5×105 huCD20-B16F10 cells were injected intravenously (i.v.) into the tail vein of C57BL/6 mice. The following day, treatments were initiated. Mice were observed daily, to monitor clinical signs. They were killed 13 days after tumor cell inoculation, and the lungs were analyzed for the presence of metastases.
Alternatively, 5×104 huCD20-B16F10 cells in a 1:1 mixture of endotoxin-free PBS and Matrigel (Corning) were injected s.c. into the flank of μMt- recipient mice (C57BL/6 background). Treatment was initiated the day after implantation and repeated on days 9 and 16.
Tumor size was monitored with a digital caliper (Mitutoyo) every three to four days and is expressed as a volume ((length× width2) /2).
For NK cell depletion, 100 μL of polyclonal anti-asialo-GM1 (Biolegend, Poly21460) antibody was injected i.p. into recipient mice two days before treatment and then once a week. Normal rabbit serum was administered as a control.
For the disseminated tumor model, 5×105 huCD20-B16F10 cells were injected intravenously (i.v.) into the tail vein of C57BL/6 mice. The following day, treatments were initiated. Mice were observed daily, to monitor clinical signs. They were killed 13 days after tumor cell inoculation, and the lungs were analyzed for the presence of metastases.
3
NK Cell Depletion in Mouse Models
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To deplete NK cells in NOD-scid mice, anti-asialo GM1 (Biolegend #146002) was i.p. injected to the recipient mice at a dose of 25 μl per mouse every 3 days. To deplete NK cells in B6-scid mice, anti-NK1.1 antibody (Bio X cell # BE0036) was given to the recipient mice at a dose of 25 μg per mouse by i.p. injection every 3 days.
4
NK Cell Depletion Protocol
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To deplete NK cells, Anti-Asialo GM1 antibodies (BioLegend, clone Poly21460) at the dose of 20 μL per mouse, were injected i.p. 1 day before i.v. injection of EMT6-Luc cells into EMT6-primed mice. Efficient depletion of NK cells were shown by flow analysis of PMBCs 3 and 5 weeks post injection of depletion antibodies. Anti-Asialo GM1 (#146007, Biolegend) and NKp46 (# 137605, Biolegend) are used to determine the NK cells in CD3-negative lymphocytes population.
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