Revision 6

#39475

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 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.

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).

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

W: 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

M: Mouse

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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).
Western Blotting Image 1: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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.
Immunoprecipitation Image 1: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb.
Immunohistochemistry Image 1: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using IL-2Rα/CD25 (E9W2J) XP® Rabbit mAb.
Immunohistochemistry Image 2: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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 3: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 4: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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).
Immunohistochemistry Image 5: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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 1: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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 2: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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.
Immunofluorescence Image 3: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
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 1: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

Cell Signaling Technology Logo
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.
Flow Cytometry Image 2: IL-2Rα/CD25 (E9W2J) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.