Application Data
Mouse anti-Rat CD25 Monoclonal Antibody | anti-CD25 antibody
Anti-Rat CD25 Biotin (Clone OX-39) (mouse IgG1)
Reactivity
Rat
Applications
Flow Cytometry, Functional Assay, Immunohistochemistry
Synonyms
CD25, Antibody; Anti-Rat CD25 Biotin (Clone OX-39) (mouse IgG1); CD25 Biotin (Clone OX-39) (mouse IgG1); Biotin Anti-Rat CD25 (IL-2R) Monoclonal Antibody; anti-CD25 antibody
Host
Mouse
Reactivity
Rat
Clonality
Monoclonal
Isotype
Mouse IgG1
Clone Number
MRC OX-39
Specificity
CD25 (IL-2Ra) (Rat CD25 (IL-2R))
Form/Format
Biotin (Biotin conjugated Ig buffered in PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. (Purified from bioreactor supernatant))
Concentration
0.1 mg/ml (varies by lot)
Sequence Length
160
Applicable Applications for anti-CD25 antibody
FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry)
Hybridoma Production
Immunization:
Immunogen:
T blasts from a mixed lymphocyte reaction between purified CD4 positive T cells and irradiated spleens.
Immunogen:
T blasts from a mixed lymphocyte reaction between purified CD4 positive T cells and irradiated spleens.
Donor
BALB/c spleen
FLOW CYTOMETRY ANALYSIS
Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-Rat cell separation medium .
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x107 cells/ml in media A. Add 50 ?l of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test).
4. To each tube, add 1.0-.5ug* of MBS520072 per 106 cells.5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 ul of secondary antibody (Streptavidin-PE) at a 1:50 dilution.
9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 ?l ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 ?l of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-Rat cell separation medium .
2. Wash 2 times.
3. Resuspend the cells to a concentration of 2x107 cells/ml in media A. Add 50 ?l of this suspension to each tube (each tube will then contain 1 x 106 cells, representing 1 test).
4. To each tube, add 1.0-.5ug* of MBS520072 per 106 cells.5. Vortex the tubes to ensure thorough mixing of antibody and cells.
6. Incubate the tubes for 30 minutes at 4°C.
7. Wash 2 times at 4°C.
8. Add 100 ul of secondary antibody (Streptavidin-PE) at a 1:50 dilution.
9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive).
10. Wash 2 times at 4°C.
11. Resuspend the cell pellet in 50 ?l ice cold media B.
12. Transfer to suitable tubes for flow cytometric analysis containing 15 ?l of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
Media
A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 ul of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ul of 2M sodium azide in 100 mls).
B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ul of 2M sodium azide in 100 mls).
Preparation and Storage
Store at 4°C. For long term storage, aliquot and freeze unused portion at -20°C in volumes appropriate for single usage.
Avoid freeze/thaw cycles.
Avoid freeze/thaw cycles.
Application Data
Application Data
Related Product Information for anti-CD25 antibody
anti-rat Interleukin-2 receptor monoclonal antibody recognizes the smaller (alpha subunit) 55kD chain of the IL-2 receptor found on activated rat T cells, thymic dendritic cells but not resting lymphocytes (1). MBS520072 binds to the rat interleukin-2 receptor and has proven to be an important marker for activated T cells (2). This clone is reported to work with frozen and paraffin sections (3).
NCBI and Uniprot Product Information
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Product Notes
The CD25 pmp22 (Catalog #AAA74138) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-Rat CD25 Biotin (Clone OX-39) (mouse IgG1) reacts with Rat and may cross-react with other species as described in the data sheet. AAA Biotech's CD25 can be used in a range of immunoassay formats including, but not limited to, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry). Researchers should empirically determine the suitability of the CD25 pmp22 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD25, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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