|H M R||Endogenous||125||Rabbit IgG|
Immunoprecipitation of extracts from K562 cells, using Jak2 (D2E12) XP® Rabbit mAb (Sepharose® Bead Conjugate) (lane 2). XP® Rabbit IgG (DA1E) mAb Isotype Control (Sepharose® Bead Conjugate) #3423 was used as a negative control (lane 1). The western blot was probed using Jak2 (D2E12) XP® Rabbit mAb #3230 and detected with Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb #3678 followed by Anti-mouse IgG, HRP-linked Antibody #7076.Learn more about how we get our images.
This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
10X Cell Lysis Buffer: (#9803) 20 mM Tris (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 2.5 mM Sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 μg/ml Leupeptin
NOTE: CST recommends adding 1 mM PMSF (#8553) before use*.
Proceed to one of the following specific set of steps.
NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127).
posted December 2007
Protocol Id: 27
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol. Store at –20°C. Do not aliquot the antibodies.
Jak2 (D2E12) XP® Rabbit mAb (Sepharose® Bead Conjugate) detects endogenous levels of total Jak2 protein. No cross-reactivity was observed with other family members.
Human, Mouse, Rat
Hamster, Monkey, Chicken, Xenopus, Bovine, Dog, Pig, Horse
Monoclonal antibody is produced by immunzing animals with a synthetic peptide corresponding to residues surrounding Pro841 of Jak2.
This Cell Signaling Technology antibody is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated Sepharose® beads. Jak2 (D2E12) XP® Rabbit mAb (Sepharose® Bead Conjugate) is useful for the immunoprecipitation of Jak2 proteins.
Members of the Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) are activated by ligands binding to a number of associated cytokine receptors (1). Upon cytokine receptor activation, Jak proteins become autophosphorylated and phosphorylate their associated receptors to provide multiple binding sites for signaling proteins. These associated signaling proteins, such as Stats (2), Shc (3), insulin receptor substrates (4), and focal adhesion kinase (FAK) (5), typically contain SH2 or other phospho-tyrosine-binding domains.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. XP is a registered trademark of Cell Signaling Technology, Inc. Jak antibodies produced under license (granting certain rights including those under U.S. Patent No. 5,658,791) from Chemicon International, Inc.
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|4089S||400 µl (40 immunoprecipitations)||$ 341.0|