Rat IL-2 Recombinant Rabbit Monoclonal Antibody [PSH05-72] - BSA and Azide free (Detector)
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Rat IL-2 aa 21-155.
Species Reactivity
Rat
Validated Applications
ELISA(Det)
Target Molecular Weight
Predicted band size: 17.6 kDa
Positive Control
Recombinant standard Rat IL-2 protein (HA210683).
Conjugation
unconjugated
Clone Number
PSH05-72
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Lot Concentration Lookup
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
ELISA(Det)
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH05-71] to Rat IL-2 (Capture) (HA722423) and recombinant standard Rat IL-2 protein (HA210683) as the standard. The reference range value is 30.9-2,500 pg/mL.
Target
Function
Cytokine produced by activated CD4-positive helper T-cells and to a lesser extend activated CD8-positive T-cells and natural killer (NK) cells that plays pivotal roles in the immune response and tolerance. Binds to a receptor complex composed of either the high-affinity trimeric IL-2R (IL2RA/CD25, IL2RB/CD122 and IL2RG/CD132) or the low-affinity dimeric IL-2R (IL2RB and IL2RG). Interaction with the receptor leads to oligomerization and conformation changes in the IL-2R subunits resulting in downstream signaling starting with phosphorylation of JAK1 and JAK3. In turn, JAK1 and JAK3 phosphorylate the receptor to form a docking site leading to the phosphorylation of several substrates including STAT5. This process leads to activation of several pathways including STAT, phosphoinositide-3-kinase/PI3K and mitogen-activated protein kinase/MAPK pathways. Functions as a T-cell growth factor and can increase NK-cell cytolytic activity as well. Promotes strong proliferation of activated B-cells and subsequently immunoglobulin production. Plays a pivotal role in regulating the adaptive immune system by controlling the survival and proliferation of regulatory T-cells, which are required for the maintenance of immune tolerance. Moreover, participates in the differentiation and homeostasis of effector T-cell subsets, including Th1, Th2, Th17 as well as memory CD8-positive T-cells.
Background References
1. Miyazaki T., Kawahara A., Fujii H., Nakagawa Y., Minami Y., Liu Z.J., Oishi I., Silvennoinen O., Witthuhn B.A., Ihle J.N. Functional activation of Jak1 and Jak3 by selective association with IL-2 receptor subunits. Science 266:1045-1047 (1994)
2. Mingari M.C., Gerosa F., Carra G., Accolla R.S., Moretta A., Zubler R.H., Waldmann T.A., Moretta L. Human interleukin-2 promotes proliferation of activated B cells via surface receptors similar to those of activated T cells. Nature 312:641-643 (1984)
Sandwich ELISA analysis of rat IL-2 matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722423) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted recombinant standard Rat IL-2 protein (HA210683) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA722423, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Interpolated concentrations of native IL-2 in Concanavalin A treated rat spleen(18 hours) supernatant samples. Rat spleen was cultured for 18 hours in the presence or absence of 3 µg/ml Concanavalin A. The concentrations of IL-2 measured in duplicate and interpolated from the IL-2 standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 8% and stimulated 8%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-2 concentration was determined to be 4,659 pg/ml in Concanavalin A stimulated rat spleen supernatant and undetectable in unstimulated rat spleen supernatant.
Interpolated concentrations of spiked IL-2 in rat cell culture media samples. The concentrations of IL-2 were interpolated from the IL-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.
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