anti-Mouse (Murine) IL5 antibody for ELISpot

Recommended IL5 Antibody (supplied by: Log in to see )

Antigen
Interleukin 5 (IL5) Antibodies
  • IL5
  • Il-5
  • IL-5
  • TRF
  • EDF
  • Pro-TRH
  • interleukin 5
  • Interleukin 5
  • interleukin 5 (colony-stimulating factor, eosinophil)
  • interleukin-5-like
  • thyrotropin-releasing hormone
  • IL5
  • Il5
  • IL-5
  • LOC100358075
  • TRH
Reactivity
Mouse (Murine)
188
137
27
20
17
3
2
2
1
1
1
Host
121
101
31
2
Conjugate
This IL5 antibody is un-conjugated
36
14
7
7
6
6
5
5
5
4
3
3
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
Application
ELISpot
123
122
107
23
22
13
11
10
9
9
9
7
6
6
5
4
4
2
2
1
1
1
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Catalog No. ABIN1177354
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Similar anti-IL5 Antibodies

Application / Reactivity Mouse (Murine)
Blocking Antibody (Inhibition) 2 Antibodies
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Cytometry by Time of Flight (CyTOF) 1 Antibodies
ELISA 57 Antibodies
ELISA (Capture) 2 Antibodies
ELISpot 1 Antibodies
Enzyme Immunoassay (EIA) 3 Antibodies
Flow Cytometry (FACS) 69 Antibodies
Functional Studies (Func) 4 Antibodies
Immunocytochemistry (ICC) 1 Antibodies
Immunofluorescence (IF) 6 Antibodies
Immunofluorescence (Paraffin-embedded Sections) (IF (p)) 13 Antibodies
Immunohistochemistry (IHC) 4 Antibodies
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) 6 Antibodies
Immunoprecipitation (IP) 2 Antibodies

General

Antigen Interleukin 5 (IL5) Antibodies
Reactivity Mouse (Murine)
(188), (137), (27), (20), (17), (3), (2), (2), (1), (1), (1)
Host
(121), (101), (31), (2)
Conjugate This IL5 antibody is un-conjugated
(36), (14), (7), (7), (6), (6), (5), (5), (5), (4), (3), (3), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application ELISpot
(123), (122), (107), (23), (22), (13), (11), (10), (9), (9), (9), (7), (6), (6), (5), (4), (4), (2), (2), (1), (1), (1)
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Product Details anti-IL5 Antibody

Target Details IL5 Application Details Handling Images
Purification Purified from tissue culture supernatant or ascites by affinity chromatography
Sterility 0.2 μm filtered
Endotoxin Level ≤0.01 ng/µg
Components 51-1805KZ: Mouse/Human IL-5 ELISPOT Capture Antibody. Storage Buffer: No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm filtered. Endotoxin level is ≤0.01 ng/μg of protein.
51-1806KZ: Mouse IL-5 ELISPOT Detection Antibody. Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.

Target Details IL5

Product Details anti-IL5 Antibody Application Details Handling Images back to top
Antigen
Alternative Name IL-5 (IL5 Antibody Abstract)
Background The enzyme-linked immunospot (ELISPOT) assay is a powerful tool for detecting and enumerating individual cells that secrete a particular protein in vitro. Based on the sandwich ELISA, the ELISPOT assay derives its specificity and sensitivity by employing high affinity capture and detection antibodies and enzyme-amplification. Although originally developed for analyzing specific antibody-secreting cells, the assay has been adapted for measuring the frequencies of cells that produce and secrete other effector molecules, such as cytokines. The sensitivity of the assay lends itself to measurement of even very low frequencies of cytokine producing cells (e.g., 1/300,000). Unique strengths of the assay include high sensitivity, high throughput, high content analysis, minimal volume of biological material required, applicability to frozen/thawed biological samples, and compatibility with other assays. For example, cells analyzed by ELISPOT can be transferred for cloning, proliferation assays, flow cytometry, or other methods of analysis. This product contains sufficient reagents for five 96-well plates, including unlabelled anti-cytokine capture antibody (no azide/low endotoxin format), biotinylated anti-cytokine detection antibody and a Certificate of Analysis that provides lot-specific optimal reagent concentrations
Research Area Cancer
Pathways JAK-STAT Signaling, Positive Regulation of Peptide Hormone Secretion, Production of Molecular Mediator of Immune Response, Feeding Behaviour

Application Details

Product Details anti-IL5 Antibody Target Details IL5 Handling Images back to top
Protocol NOTE: Use ELISPOT plates and reagents under aseptic conditions (e.g., using a laminar flow hood or biosafety cabinet) for Steps 1-7.

Coating Antibody
1. Dilute capture antibody in Coating Buffer (see Certificate of Analysis for antibody dilution information). Add 100 µl of diluted antibody solution to each well of an ELISPOT plate, Millipore plate, Cat. No. S2EM004M99 is recommended.
2. Store plates at 4°C overnight with lid.

Blocking
3. Discard Coating Antibody. Wash wells 1X with 200 µL/well Blocking Solution.
4. Add 200 µL/well Blocking Solution and incubate for 2 hr at room temperature.

Cell Activation
5. Discard Blocking Solution. Prepare mitogen or antigen, diluted in complete tissue culture medium (e.g., RPMI 1640 with FBS, Pen/Strep, and L-glutamine). Add 100 µL/well to ELISPOT plate.
6. Prepare cell suspensions at different densities, (e.g., 1 X 10e5/mL - 2 X 10e6 cells/mL) Add 100 µL/well of each cell suspension to ELISPOT plate wells.
7. Replace ELISPOT plate lid. Incubate ELISPOT plate at 37°C, 5% CO2 and 99% humidity. The duration of the incubation time can be varied (e.g., 2 hr - 24 hr) depending on the nature of the stimulatory cell culture system. Specific activation conditions and incubation times will vary, depending on cell type, kinetics, and cytokine of interest. Please see Certificate of Analysis provided with each ELISPOT Pair for assay conditions of suggested positive controls. Note: After step 7, aseptic conditions are no longer needed.

Detection Antibody
8. Aspirate cell suspension. Wash wells 2X with deionized (DI) water. Allow wells to soak for 3-5 min at each wash step.
9. Wash wells 3X with 200 µL/well Wash Buffer I. Discard Wash Buffer.
10. Dilute Detection Antibody in Dilution Buffer (see Certificate of Analysis for antibody dilution information). Add 100 µL per well.
11. Replace lid and incubate for 2 hr at room temperature.

Enzyme
12. Discard Detection Antibody solution. Wash wells 3X with 200 µL/well Wash Buffer I.
13. Dilute Streptavidin-Horseradish Peroxidase (HRP) in Dilution Buffer. (BD™ ELISPOT Streptavidin-HRP, Cat. No. 557630 is recommended). Add 100 µL/well diluted Streptavidin-HRP.
14. Replace lid, incubate for 1 hr at room temperature.

Substrate
15. Discard Streptavidin-HRP solution. Wash wells 4X with 200 µL/well Wash Buffer I.
16. Wash wells 2X with 200 µL/well Wash Buffer II.
17. Add 100 µL of Final Substrate Solution (AEC) to each well. Monitor spot development from 5 - 60 min.
18. Stop substrate reaction by washing wells with DI water.
19. Air-dry plate for 2 hr - overnight at room temperature in the dark, until the plate is completely dry. Store plate in the dark, prior to analysis.
20. Enumerate spots manually using a dissecting microscope or automatically using an ELISPOT Analyzer.
Restrictions For Research Use only

Handling

Product Details anti-IL5 Antibody Target Details IL5 Application Details Images back to top
Format Liquid
Buffer Aqueous buffered solution
Storage 4 °C