Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-F, FC-FP, FC-L

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

40-80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P01590

Entrez-Gene Id:

16184

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:50 - 1:200
Immunofluorescence (Frozen) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
Flow Cytometry (Live) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #95742.

Specificity / Sensitivity

IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb recognizes endogenous levels of total IL-2Rα/CD25 protein. This antibody non-specifically labels myenteric nerves in the gut.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of mouse IL-2Rα/CD25 protein.

Background

Interleukin-2 (IL-2) is a T cell stimulatory cytokine best known for inducing T cell proliferation and NK cell proliferation and activation (1,2). IL-2 also promotes peripheral development of regulatory T cells (Tregs) (3,4). Conversely, IL-2 is involved in the activation-induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (5). The effects of IL-2 are mediated through a trimeric receptor complex consisting of IL-2Rα, IL-2Rβ, and the common gamma chain, γc (1,2). IL-2Rα binds exclusively to IL-2 with low affinity and increases the binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-15 also binds to IL-2Rβ (1,2). γc is used by other cytokines, including IL-4, IL-7, IL-9, IL-15, and IL-21 (1,2). Binding of IL-2 initiates signaling cascades involving Jak1, Jak3, Stat5, and the PI3K/Akt pathways (1,2).

  1. Ma, A. et al. (2006) Annu Rev Immunol 24, 657-79.
  2. Gaffen, S.L. and Liu, K.D. (2004) Cytokine 28, 109-23.
  3. Fehérvari, Z. et al. (2006) Trends Immunol 27, 109-11.
  4. Antony, P.A. et al. (2006) J Immunol 176, 5255-66.
  5. Jaleco, S. et al. (2003) J Immunol 171, 61-8.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) FC-FP: Flow Cytometry (Fixed/Permeabilized) FC-L: Flow Cytometry (Live)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 6
#39475

IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb

Western Blotting Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cells using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunoprecipitation of IL-2Rα/CD25 protein from CTLL-2 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb. Western blot analysis was performed using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunohistochemistry Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse thymus using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb.
Immunohistochemistry Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb.
Immunohistochemistry Image 3: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded CTLL-2 cell pellet (left, positive) or NIH/3T3 cell pellet (right, negative) using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb.
Immunohistochemistry Image 4: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse colon using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (left) or IL-2Rα/CD25 (E4B2Z) XP® Rabbit mAb #36128 (right). These two antibodies detect independent, unique epitopes on mouse IL-2Rα/CD25. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 5: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumor in mouse lung using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunofluorescence Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse spleen using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled with CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127 (blue pseudocolor) and FoxP3 (D6O8R) Rabbit mAb (Alexa Fluor® 647 Conjugate) (red).
Immunofluorescence Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse spleen using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (green), CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127 (red pseudocolor), and CD45R/B220 (RA3-6B2) Rat mAb (redFluor 710 Conjugate) #82984 (cyan pseudocolor). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse colon using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (green) and CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127 (red pseudocolor). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue). Note: signal in myenteric plexus is presumed to be non-specific.
Flow Cytometry Image 1: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Flow cytometric analysis of live mouse splenocytes using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (right) or concentration- matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), and co-stained with CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
Flow Cytometry Image 2: IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb Expand Image
Flow cytometric analysis of live C2C12 cells (blue, negative) and CTLL-2 cells (green, positive) using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.