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Mouse (Murine) STAT6 ELISA Kit

Recommended STAT6 Product (supplied by: Log in to see )

Antigen
Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced (STAT6) ELISA Kits
  • D12S1644
  • IL-4-STAT
  • STAT6B
  • STAT6C
Epitope
pTyr641
Reactivity
Human, Mouse (Murine)
Alternatives
Kits with alternative reactivity to:
14
12
10
3
3
2
2
2
2
1
1
1
Methode Type
Cell ELISA
Application
ELISA
Supplier
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Catalog No. ABIN1981845
$ 446.60
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
0.0012121415 ABIN773786 Competition ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Body Fluids Log in to see
0.0012121415 ABIN1117118 Sandwich ELISA Serum, Plasma, Biological Fluids Log in to see
0.0012121415 ABIN1380711 Cell ELISA Log in to see
0.0012121415 ABIN1380710 Cell ELISA Log in to see
0.0012121415 ABIN510875 Serum, Plasma, Cell Culture Supernatant, Body Fluids, Tissue Homogenate Log in to see
0.0012121415 ABIN2114938 Cell ELISA Log in to see
0.0012121415 ABIN2114937 Cell ELISA Log in to see
0.0012121415 ABIN2866207 Cell ELISA Cell Culture Cells Log in to see
0.0012121415 ABIN2866206 Cell ELISA Cell Culture Cells Log in to see
0.0012121415 ABIN1158102 Log in to see
0.0012121415 ABIN2105722 Sandwich ELISA Log in to see
0.0012121415 ABIN2951430 78.125-5000 pg/mL 78.125 pg/mL Log in to see

Top referenced STAT6 ELISA Kit for ELISA

  • Human STAT6 ELISA Kit for Cell ELISA - ABIN1981845 (8 Publications): Quelle, Shimoda, Thierfelder, Fischer, Kim, Ruben, Cleveland, Pierce, Keegan, Nelms: Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis. in Molecular and cellular biology 1995 (PubMed)

Similar STAT6 ELISA Kits

Application / Reactivity Human Mouse (Murine)
ELISA 15 ELISA Kits 13 ELISA Kits

General

Antigen Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced (STAT6) ELISA Kits
Epitope pTyr641
Reactivity Human, Mouse (Murine)
Kits with alternative reactivity to:
(14), (12), (10), (3), (3), (2), (2), (2), (2), (1), (1), (1)
Methode Type Cell ELISA
Application ELISA
Pubmed 8 references available
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Product Details STAT6 ELISA Kit

Target details Application Details Handling References for STAT6 Kit (ABIN1981845) Images
Purpose Semi-Quantitative Cell-Based Human/Mouse Stat 6 (Tyr641) Phosphorylation ELISA Kit for adherent whole cell lines.
Brand CellBIND®
Sample Type Adherent Cell Culture
Detection Method Colorimetric
Characteristics Protein phosphorylation is instrumental in the regulation of protein activity within a cell. It plays important roles in the living cells including proliferation, differentiation and metabolism. A large number of protein kinases and phosphatases have been extensively investigated, and have been shown to be involved in signal transduction pathways.
The Cell-Based Human/Mouse STAT6 (Tyr641) Phosphorylation ELISA Kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of STAT6 (Tyr641) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured human and mouse cell lines. By determining STAT6 protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot. In the Cell-Based STAT6 (Tyr641) Phosphorylation ELISA kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, an antiphospho- STAT6 (Tyr641) or anti-STAT6 antibody is pipetted into the wells and incubated. The wells are washed and an HRP-conjugated antimouse IgG is added to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Components A, Uncoated 96-Well Microplate, 1 plate, Room Temperature
B, 20X Wash Buffer A Concentrate, 1 vial (30 ml), 2-8 °C
C, 20X Wash Buffer B Concentrate, 1 vial (30 ml), 2-8 °C
D, Fixing Solution, 1 vial (30 ml), 2-8 °C
E, 30X Quenching Buffer Concentrate, 1 vial (2 ml), 2-8 °C
F, 5X Blocking Buffer Concentrate, 1 vial (20 ml), 2-8 °C (1 month) G, 500X Rabbit Anti-phospho (Tyr641) STAT6 Concentrate, 1 vial (10 μl), -20 °C
H, 5000X Mouse Anti-STAT6 Concentrate, 1 vial (5 µL ), -20 °C
I-1, 1000X HRP Conjugated Anti-Rabbit IgG Concentrate, 1 vial (10 µL ), -20 °C
I-2, 1000X HRP Conjugated Anti-Mouse IgG Concentrate, 1 vial (10 µL ), -20 °C
J, TMB Substrate, 1 vial (12 ml), 2-8 °C
K, Stop Solution** 1 vial (14 ml), 2-8 °C

*For up to 3 months (unless otherwise stated) or until expiration date.
**Contains 0.2 M Sulfuric Acid
Material not included 1. A model cell line, protein tyrosine kinase inhibitors, growth factors or cytokines
2. Microplate reader capable of measuring absorbance at 450 nm
3. 37 °C incubator
4. Precision pipettes to deliver 2 µL to 1 ml volumes
5. Adjustable 1-25 ml pipettes for reagent preparation
6. 100 ml and 1 liter graduated cylinders
7. Absorbent paper
8. Distilled or deionized water
9. Orbital shaker or oscillating rocker

Target details

Product Details STAT6 ELISA Kit Application Details Handling References for STAT6 Kit (ABIN1981845) Images back to top
Antigen
Alternative Name STAT6 (STAT6 ELISA Kit Abstract)
Research Area Chromatin and Nuclear Signaling, Signaling, Receptors, Inflammation
Pathways JAK-STAT Signaling

Application Details

Product Details STAT6 ELISA Kit Target details Handling References for STAT6 Kit (ABIN1981845) Images back to top
Comment

Quality control: Representative results of Cell-Based STAT6 (Tyr641) are shown below:
1. Seeded 100 µL of 30,000 A431 cells into appropriate wells of the microplate. Cells were incubated at 37 °C in 5 % CO2 overnight.
2. Added 50 µL of different concentrations of stimulators (rhEGF concentration for A431 cells: 0, 20 or 100 ng/mL in serum free DMEM) to appropriate wells (shown below). Then incubated for 10, 20, or 30 min at 37 °C.
3. Discarded the solution and washed 3 times with 1X Wash Buffer A (200 µL each) immediately. Then flipped the microplate upside down and gently tapped to remove all of excess wash buffer. The protocol was continued as stated.

Plate 96 wells
Protocol 1. Add cells
2. Treatment with stimulators or inhibitors
3. Fixing and blocking
4. Anti-phospho-protein antibody or anti-pan-protein antibody
5. HRP-conjugated secondary antibody
6. Develop with substrate
Reagent Preparation NOTE: Thaw all reagents to room temperature immediately before use. If wash buffers contain visible crystals, warm to room temperature and mix gently until dissolved.
NOTE: Briefly centrifuge (~1,000g) ITEMS G, H, and I before opening to ensure maximum recovery.
For more information look at the picture.
Assay Procedure NOTE: ALL incubations and wash steps must be performed under gentle rocking or rotation (~1-2 cycles/sec).
1. Design your experiment. For example, see Figure 2 below.
OPTIONAL: If seeding HUVECs, HMEC-1 or other loosely attached cells, coat the Uncoated 96-Well Microplate (ITEM A) by adding 100 µL poly-L-Lysine (Recommended Sigma Aldrich, Cat#: P4832) into each well and then follow manufacturer's instructions. A pre-coated CellBIND® microplate or other poly-lysine treated tissue culture plate may be used in place of Item A.
2. Seed 100 µL of 30,000 cells into each well of the Uncoated 96-Well Microplate (ITEM A) provided and incubate overnight at 37 °C with 5 % CO2.
NOTE: The optimal cell number used will vary on the cell line and the relative amount of protein phosphorylation. More or less cells may be used but this must be determined empirically.
NOTE: The cells can be starved ~4-24 hours (depending on cell line) prior to treatment with inhibitors or activators.
3. Apply various treatments, inhibitors (such as siRNA or chemicals) or activators according to manufacturer's instructions and incubate for the desired time points.
NOTE: It is recommended to dissolve inhibitors or activators into serum-free cell culture medium before treating the cells (unless otherwise stated in the manufacturer's instructions.)
4. Discard the cell culture medium by flipping the microplate upside down and gently tapping the bottom of the microplate over a sink.
5. Wash by pipetting 200 µL of the prepared 1X Wash Buffer A (ITEM B) into each well. Discard the wash buffer (same as step 4) and wash 2 more times for a total of 3 washes using fresh wash buffer each time. After the final wash, gently blot the microplate onto a paper towel to remove any excess/remaining buffer.
NOTE: To avoid cell loss, do not pipette directly onto the cells. Instead, gently dispense the liquid down the wall of cell culture wells. Also avoid the use of vacuum suction or too forcefully tapping the microplate when discarding any solution.
6. Add 100 µL of Fixing Solution (ITEM D) into each well and incubate for 20 minutes at room temperature.
NOTE: The fixing solution is used to permeabilize the cells.
7. Repeat wash step 5.
8. Add 200 µL of the prepared 1X Quenching Buffer (ITEM E) into each well and incubate 20 minutes at room temperature.
NOTE: The quenching buffer is used to minimize the background response.
9. Wash 4 times with 1X Wash Buffer A.
10. Add 200 µL of the prepared 1X Blocking Buffer (ITEM F) into each well and incubate for 1 hour at 37 °C.
11. Wash 3 times with the prepared 1X Wash Buffer B (ITEM C).
NOTE: If needed, the microplate may be stored at -80 °C for several days after this wash.
12. Add 50 µL of the prepared 1X primary antibody (ITEM G or H) into each corresponding well and incubate for 2 hours at room temperature.
13. Wash 4 times with 1X Wash Buffer B.
14. Add 50 µL of the prepared 1X HRP Conjugated secondary antibody (ITEM I-1 or I-2) into each well and incubate for 1 hour at room temperature.
NOTE: Item I-1 is the secondary antibody for Item G (primary antibody). Item I-2 is the secondary antibody for Item H (primary antibody).
15. Wash 4 times with 1X Wash Buffer B.
16. Add 100 µL of the TMB Substrate (ITEM J) into each well and incubate for 30 minutes at room temperature in the dark.
17. Add 50 µL of the Stop Solution (ITEM K) into each well. Read at 450 nm immediately.
Restrictions For Research Use only

Handling

Product Details STAT6 ELISA Kit Target details Application Details References for STAT6 Kit (ABIN1981845) Images back to top
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment Store entire kit at ≤ -20 °C immediately upon arrival.
Expiry Date 6 months

References for STAT6 Kit (ABIN1981845)

Product Details STAT6 ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Patel, Wang, Lee, Taylor, Pierce, LaRochelle: "Stat6 and Jak1 are common elements in platelet-derived growth factor and interleukin-4 signal transduction pathways in NIH 3T3 fibroblasts." in: The Journal of biological chemistry, Vol. 271, Issue 36, pp. 22175-82, 1996

Patel, Wang, Lee, Taylor, Pierce, LaRochelle: "Stat6 and Jak1 are common elements in platelet-derived growth factor and interleukin-4 signal transduction pathways in NIH 3T3 fibroblasts." in: The Journal of biological chemistry, Vol. 271, Issue 36, pp. 22175-82, 1996

Patel, Wang, Lee, Taylor, Pierce, LaRochelle: "Stat6 and Jak1 are common elements in platelet-derived growth factor and interleukin-4 signal transduction pathways in NIH 3T3 fibroblasts." in: The Journal of biological chemistry, Vol. 271, Issue 36, pp. 22175-82, 1996

Quelle, Shimoda, Thierfelder, Fischer, Kim, Ruben, Cleveland, Pierce, Keegan, Nelms: "Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis." in: Molecular and cellular biology, Vol. 15, Issue 6, pp. 3336-43, 1995

Quelle, Shimoda, Thierfelder, Fischer, Kim, Ruben, Cleveland, Pierce, Keegan, Nelms: "Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis." in: Molecular and cellular biology, Vol. 15, Issue 6, pp. 3336-43, 1995

Quelle, Shimoda, Thierfelder, Fischer, Kim, Ruben, Cleveland, Pierce, Keegan, Nelms: "Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis." in: Molecular and cellular biology, Vol. 15, Issue 6, pp. 3336-43, 1995

Background publications

Patel, Wang, Lee, Taylor, Pierce, LaRochelle: "Stat6 and Jak1 are common elements in platelet-derived growth factor and interleukin-4 signal transduction pathways in NIH 3T3 fibroblasts." in: The Journal of biological chemistry, Vol. 271, Issue 36, pp. 22175-82, 1996

Quelle, Shimoda, Thierfelder, Fischer, Kim, Ruben, Cleveland, Pierce, Keegan, Nelms: "Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis." in: Molecular and cellular biology, Vol. 15, Issue 6, pp. 3336-43, 1995

Images

Product Details STAT6 ELISA Kit Target details Application Details Handling References for STAT6 Kit (ABIN1981845) back to top
Supplier Images
 image for STAT6 ELISA Kit (Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced) (ABIN1981845) Cell-Based protein phosphorylation procedure
 image for STAT6 ELISA Kit (Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced) (ABIN1981845) This picture shows the reagent preparation.
 image for STAT6 ELISA Kit (Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced) (ABIN1981845) Example of how to seed cells for cell-based assay
 image for STAT6 ELISA Kit (Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced) (ABIN1981845) A431 cells were stimulated by different concentration of recombinant human EGF for 10...
Western Blotting (WB) image for STAT6 ELISA Kit (Signal Transducer and Activator of Transcription 6, Interleukin-4 Induced) (ABIN1981845) Western blot analysis of extracts from 100 ng/mL hEGF treated A431 cells. Phospho-Sta...