Skip to main content Skip to navigation
Polyclonal Rabbit Anti-JNK (pT183/pY185)
Polyclonal Rabbit Anti-JNK (pT183/pY185)
Analysis of JNK (pT183/pY185) in activated human lymphocytes.  Fresh blood was either stimulated with PMA and Ionomycin at 37˚C for 10 minutes (open histogram) or unstimulated (shaded histogram).  The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer (Cat. No. 558049) and permeabilized in BD Phosflow Perm Buffer III (Cat. No. 558050).  Then they were stained with Purified Rabbit anti-JNK (pT183/pY185) followed by PE F(ab')2 Donkey anti-Rabbit IgG (Cat. no. 558416).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.  For data analysis, the lymphocytes were selected by their scatter profile.
Polyclonal Rabbit Anti-JNK (pT183/pY185)

JNK (pT183/pY185) staining on tonsil.  Fresh human tonsil was incubated in 5 mM Pervanadate solution for 2 hours (top row) or untreated (bottom row), then fixed in formalin and processed.  Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the sections were either left untreated (left column) or treated with a phosphatase to eliminate all phosphorylation (right column).  The tissue sections were stained with Purified Rabbit anti-JNK (pT183/pY185) with Hematoxylin counterstaining.  Original magnification: 20X.

Analysis of JNK (pT183/pY185) in activated human lymphocytes.  Fresh blood was either stimulated with PMA and Ionomycin at 37˚C for 10 minutes (open histogram) or unstimulated (shaded histogram).  The cells were lysed and fixed with 1X BD Phosflow Lyse/Fix Buffer (Cat. No. 558049) and permeabilized in BD Phosflow Perm Buffer III (Cat. No. 558050).  Then they were stained with Purified Rabbit anti-JNK (pT183/pY185) followed by PE F(ab')2 Donkey anti-Rabbit IgG (Cat. no. 558416).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.  For data analysis, the lymphocytes were selected by their scatter profile.

JNK (pT183/pY185) staining on tonsil.  Fresh human tonsil was incubated in 5 mM Pervanadate solution for 2 hours (top row) or untreated (bottom row), then fixed in formalin and processed.  Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the sections were either left untreated (left column) or treated with a phosphatase to eliminate all phosphorylation (right column).  The tissue sections were stained with Purified Rabbit anti-JNK (pT183/pY185) with Hematoxylin counterstaining.  Original magnification: 20X.

Product Details
Down Arrow Up Arrow


BD Phosflow™
Rabbit IgG
Phosphorylated Human JNK
Intracellular staining (flow cytometry) (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required), Western blot (Tested During Development)
20 µl
AB_647297
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

The polyclonal antibody was purified from antiserum by negative adsorption and affinity chromatography. Store undiluted at 4°C.

Product Notices

  1. 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).
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
558268 Rev. 3
Antibody Details
Down Arrow Up Arrow
poly/JNK/44682

C-Jun NH2-terminal Kinases (JNK), also called Stress Activated protein Kinases (SAPK), are mitogen activated protein kinases (MAPK).  JNK/SAPK, along with p38 family and ERK, comprise three major classes of MAPK.  The JNK play a role in signal transduction in response to cytokines and various forms of environmental stress.  JNK are critical to multiple signal transduction pathways that regulate cell growth, apoptosis, and the cellular response to stress.  Complete activation of JNK/SAPK by MKK4 and MKK7 requires phosphorylation of both Threonine 183 (T183) and Tyrosine 185 (Y185), which are located in a Thr-X-Tyr motif.  Active JNK binds to c-Jun and phosphorylates it on Serine 63 and Serine 73, thus inducing expression of genes, such as proinflammatory cytokines.  Of the three isoforms; JNK1 and JNK2 are broadly expressed, and JNK3 appears primarily on neuronal cell types.

The affinity purified rabbit anti-JNK (pT183/pY185) polyclonal antibody recognizes human JNK1 and 2 phosphorylated at T183 and Y185 in flow cytometric analysis of human blood cells.  The same polyclonal antibody preparation is able to identify human and rat JNK1 and 2 (pT183/pY185) in western blot analysis.  The sequence is conserved in JNK3 and in several other species, including mouse, chicken, nematode, and fruit fly.  We recommend purified JNK (pT183/pY185) monoclonal antibody clone 41 (Cat. No. 612540 or 612541) for western blot analysis.

558268 Rev. 3
Format Details
Down Arrow Up Arrow
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
558268 Rev.3
Citations & References
Down Arrow Up Arrow

Development References (2)

  1. Fleming Y, Armstrong CG, Morrice N, Paterson A, Goedert M, Cohen P. Synergistic activation of stress-activated protein kinase 1/c-Jun N-terminal kinase (SAPK1/JNK) isoforms by mitogen-activated protein kinase kinase 4 (MKK4) and MKK7. Biochem J. 2000; 352:145-154. (Biology). View Reference
  2. Kyriakis JM, Avruch J. Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev. 2001; 81(2):807-869. (Biology). View Reference
558268 Rev. 3

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.