Anti Asialo GM1 Antibodies
The glycolipid asialo ganglioside-GM1 (ASGM1) is expressed in T cells infected with viruses and in natural killer (NK) cells. Anti asialo GM1 antibody reduces natural killer (NK) activity in cells of various strains of mice and rats.
Fujifilm Wako offers "Anti asialo GM1 (Rabbit)" which is rabbit polyclonal antibody, raised against asialo GM1. It reacts with mouse and rat natural killer cells, mouse monocytes and fetal thymocytes and is tested the anti-NK cells activity in vivo.
What is asialo GM1?
The glycolipid asialo ganglioside-GM1 (ASGM1) is expressed in T cells infected with viruses and in natural killer (NK) cells. ASGM1 is localized in lipid raft structures in NK and CD8 (+) T cells. Anti asialo GM1 antibody reduces natural killer (NK) activity in cells of various strains of mice and rats. To investigate the function of NK cells, or transplant tumor tissue derived from the other species to mice, Anti asialo GM1 antibody is injected to remove NK cells1-2). Recently, it is reported that ASGM1 is also expressed by Basophils, and Anti asialo GM1 antibody is able to remove Basophils3).
Anti asialo GM1 (Rabbit)
Anti asialo GM1 (Rabbit) is rabbit polyclonal antibody, raised against asialo GM1. It reacts with mouse and rat natural killer cells, mouse monocytes and fetal thymocytes and is tested the anti-NK cells activity in vivo.
Antibody Information
| Contents | Rabbit anti-asialo GM1, lyophilized |
|---|---|
| Preparation | GM1 from bovine brain tissue was repeatedly immunized with methylated bovine serum albumin and with complete Freund’s adjuvant. |
| Purification | Gammaglobulin fraction of serum was obtained by 50 % ammonium sulfate precipitation methods followed by dialyzing with phosphate buffered saline (pH 7.2). |
| Specificity | React with mouse and rat natural killer cells, mouse monocytes and fetal thymocytes. |
| Reconstitution | Reconstitute with 1 mL of distilled water. For dilution, phosphate buffer (pH 7.2) is recommended. |
| Injection | Inject to mouse 10-50 μL intravenously. |
Application Data
Evaluation of cytotoxic activity against murine lymphoma Yac-1 cells
Each antibody was reconstituted in 1 mL of distilled water and diluted 2-fold in RPMI 1640 medium. For NK cells activity, 200 µL of anti-asialo GM1 (Product No. 986-10001 or Product No.014–09801) antibodies were administered intraperitoneally. Two days after administration of anti-asialo GM1 antibody, the control group (Group No. 2) and the antibody administration group (Group No. 3 and 4) were administered 100 μg of Poly I:C intraperitoneally.
Spleens were collected 16 to 18 hours after polyI:C administration. Cells were prepared from the collected spleens and cytotoxicity (NK cells activity) was measured against CFDA-labeled Yac-1 cells using flow cytometry.
The evaluation was conducted under the following two conditions.
Target cells (Yac-1 cells): effector cells (spleen cells) = 1:100 or 1:50
| No. | Group | Administration (i.p.) | |
|---|---|---|---|
| 1 | Negative control | Vehicle (RPMI 1640) |
Vehicle (RPMI 1640) |
| 2 | Control | Poly I:C (100 μg/mouse) |
|
| 3 | Antibody | pAb (Product No. 986-10001) |
|
| 4 | pAb (Product No. 014-09801) |
||
[Result]
In the anti-asialo GM1 antibody administration groups, NK cells activity was significantly lower than in the comparison control under both conditions. There was no significant difference in NK activity between Group No.3 and No.4.
Evaluation of NK cell removal
The number of NK cells in the spleen collected from the mice was measured by immunostaining.
The spleen cells were stained with CD335 (NKp46) and CD3ε. CD335-positive/CD3ε-negative cells were detected as NK cells. The flow cytometer analysis diagram shows one individual from the control group and one individual from antibody administration group.
| No. | Group | Administration (i.p.) | |
|---|---|---|---|
| 1 | Negative control | Vehicle (RPMI 1640) |
Vehicle (RPMI 1640) |
| 2 | Control | Poly I:C (100 μg/mouse) |
|
| 3 | Antibody | pAb (Product No. 986-10001) |
|
| 4 | pAb (Product No. 014-09801) |
||
[Result]
Compared to the percentage of NK cells in the control group (Group No. 2), the percentage of NK cells in the antibody-treated group was significantly lower. There was no significant difference in the percentage of NK cells between Group No. 3 and 4.
The ability of the two types of anti-GM1 antibodies to remove NK cells is considered equivalent.
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Anti asialo GM1 (Rabbit)
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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