mAbs Against MOUSE Protein
CD20
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | Rat monoclonal |
| CLONE NAME: | GOT214A |
| PROTEIN: | mouse CD20 (B-lymphocyte antigen CD20/Ms4a1) |
| PROTEIN WEB: | https://www.uniprot.org/uniprot/P19437 |
| ANTIGEN USED: | RBL-1-mCD20 transfected cells |
| FUSION PARTNER: | myeloma p3-NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG2a |
| SPECIES REACTIVITY: | mouse |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
| COMMERCIALIZED BY: | Abcam |
Description
B-lymphocyte-specific membrane protein that plays a role in the regulation of cellular calcium influx necessary for the development, differentiation, and activation of B-lymphocytes. Functions as a store-operated calcium (SOC) channel component promoting calcium influx after activation by the B-cell receptor/BCR.
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GOT214A | neat | supernatant | / | membrane | ||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GOT214A | 1:15 | supernatant | Heat High pH9 at 95ºC 20 min | lymph node / Testis | membrane | |||
| Immunofluorescence | |||||||||
Enlarge image
- GOT214A antibody in HEK293 transfected cells
- To confirm that GOT214A mAb recognizes mouse CD20 protein, immunocytochemistry on frozen cytospins preparation of mouse CD20 expressed in HEK293T was performed. Anti-MYC Ab was used as a positive control. Cytospin preparation of mouse NGFR transfected cells was used as negative control.
Enlarge image
- GOT214A mAb can be used to detect mouse CD20 protein in mouse paraffin tissues
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | GOT214A | Neat | supernatant | spleen / | Available | |||
| Immunoprecipitation | ||||||||
Enlarge image
- GOT214A mAb is able to detect mouse CD20 protein by WB.
- Predicted molecular weight: 32kDa
Observed molecular weight: 32kDa
LANES
Lane 1 rat RBL-1 cell line (80ug) (-)
Lane 2 Mouse spleen extract (80ug) (+)
Lane 3 Mouse spleen extract (100ug) (-)
| FLOW CYTOMETRY | Clone | Dilution | +/- control | Type of fluorocrom | Protocol |
|---|---|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | GOT214A | 1:400 purified antibody | mouse spleen / |
Enlarge image
- GOT214A mAb can be used to detect mouse CD20 protein by flow cytometry
- Double immunolabelling with mCD3 and mIgM were used to test GOT214A clone.