Alexa Fluor® 647 Mouse Anti-ERK1/2 (pT202/pY204)

BD Phosflow™ Alexa Fluor® 647 Mouse Anti-ERK1/2 (pT202/pY204)

Name: Alexa Fluor® 647 Mouse Anti-ERK1/2 (pT202/pY204)
Brand: BD Phosflow™
SKU: 561992

クローン 20A

(RUO)

Flow cytometric analysis of ERK1/2 (pT202/pY204). Human peripheral blood mononulcear cells (PBMC) were either left unstimulated (unshaded) or stimulated (shaded) with 40 nM PMA for 10 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeabilized by adding BD Phosflow™ Perm Buffer III (Cat. No. 558050) for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with the Alexa Fluor® 647 mouse anti-ERK1/2 (pT202/pY204) antibody. Cells were analyzed on a BD FACSCalibur™ flow cytometry instrument. For intracellular staining of human whole blood, BD™ Phosflow Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.

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製品詳細

Brand: BD Phosflow™

Alternative Name: p44/42 MAPK; Extracellular signal-Regulated Kinase 1/2 (pT202/Y204)

Reactivity: Human (QC Testing), Mouse,Rat (Tested in Development)

Isotype: Mouse IgG1

Immunogen: Phosphorylated Rat ERK1 (T202/Y204) Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

RRID: AB_10896138

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

推奨アッセイ手順

For more information about BD Phosflow™: Please refer to http://www.bdbiosciences.com/support/resources/phosflow/index.jsp

Investigators may also find the following protocols to be helpful:

Phosflow protocol for human PBMC: Please refer to http://www.bdbiosciences.com/documents/Phosflow_Protocol_for_Human_PBMCs.pdf

Phosflow protocol for human whole blood: Please refer to http://www.bdbiosciences.com/documents/Phosflow_Protocol_for_Human_Whole_Blood_Samples.pdf

Phosflow protocol for adherent cells: Please refer to http://www.bdbiosciences.com/support/resources/protocols/protocol_adherent.jsp

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.

関連製品

Stain Buffer (FBS) RUO

サイズ 500 mL カタログ番号 554656

Lyse/Fix Buffer 5X RUO

サイズ 250 mL カタログ番号 558049

Fixation Buffer RUO

サイズ 100 mL カタログ番号 554655

Fix Buffer I RUO

サイズ 250 mL カタログ番号 557870

Perm Buffer III RUO

サイズ 125 mL カタログ番号 558050

Purified Mouse Anti-ERK1/2 (pT202/pY204) RUO

サイズ 50 µg カタログ番号 612358

詳細を表示

561992 Rev. 1

抗体の詳細

20A

The members of the Mitogen-Activated Protein Kinase (MAPK) family are components of a key signal transduction cascade that links events at the cell surface to responses in the nucleus. The signaling cascade is found in species as varied as yeast and humans, with many of the proteins being well conserved. In mammals the most widely studied members of the cascade are the Extracellular signal-Regulated Kinases, ERK1 (p44 MAPK) and ERK2 (p42 MAPK). ERK1 and ERK2 share 85% homology and are activated by extracellular signals such as growth factors, hormones, and phorbol esters. Activation occurs through a series of phosphorylations by kinases activating other kinases and eventually leading to phosphorylation of the ERKs. Growth factor stimulation leads to activation of Ras and Raf, leading to phosphorylation of MEK1 (MAPK/ERK kinase) which, in turn, activates the ERKs via dual phosphorylation. Once activated, the ERKs phosphorylate other cytoplasmic signalling molecules, cell-surface receptors, microtubule-associated proteins, and transcription factors in the nucleus. Thus, the active ERK has myriad downstream effectors that implicate it in the control of cell proliferation and differentiation, as well as regulation of the cytoskeleton. Furthermore, studies have shown that elevated ERK activity is associated with some cancers.

The 20A monoclonal antibody recognizes the phosphorylated threonine 202 and tyrosine 204 (pT202/pY204) of human ERK1 and pT184/pY186 of human ERK2. The orthologous phosphorylation sites in murine ERK1 and ERK2 are T203/Y205 and T183/Y185.

561992 Rev. 1

フォーマットの詳細

Alexa Fluor™ 647

Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

フォーマット: Alexa Fluor™ 647

励起源: Red 627-640 nm

最大励起波長: 653 nm

最大蛍光波長: 669 nm

561992 Rev.1

引用＆参考文献

Development References (3)

Boulton TG, Cobb MH. Identification of multiple extracellular signal-regulated kinases (ERKs) with antipeptide antibodies. Cell Regul. 1991; 2(5):357-371. (Biology). View Reference
Clark EA, Hynes RO. Ras activation is necessary for integrin-mediated activation of extracellular signal-regulated kinase 2 and cytosolic phospholipase A2 but not for cytoskeletal organization. J Biol Chem. 1996; 271(25):14814-14818. (Biology). View Reference
Sivaraman VS, Wang H, Nuovo GJ, Malbon CC. Hyperexpression of mitogen-activated protein kinase in human breast cancer. J Clin Invest. 1997; 99(7):1478-1483. (Biology). View Reference

561992 Rev. 1

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.