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89892
Phospho-Tau (Ser214/T217) Signaling Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Phospho-Tau (Ser214/T217) Signaling Antibody Sampler Kit #89892

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Confocal immunofluorescent analysis of fixed frozen mouse spleen labeled with p70 S6 Kinase (E8K6T)XP(R) Rabbit mAb (left, green) and colabeled with CD45R/B220 (RA3-6B2) Rat mAb (redFluor 710 Conjugate) #82984 (center, red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse kidney, labeled with p70 S6 Kinase (E8K6T)XP® Rabbit mAb (left, green) and DyLight 554 Phalloidin #13054 (right, red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of MCF7 cells, wild-type (left, positive) or p70 S6 Kinase-1/ p70 S6 Kinase-2 double knockout (right, negative) using p70 S6 Kinase (E6L9T) XP® Rabbit mAb (green) and DAPI #4083 (blue). Cell lines courtesy of Dr. Mario Pende, INSERM, Paris, France.
Simple Western analysis of lysates (0.1 mg/mL) from Mouse Brain Tissue Extracts using Tau (D1M9X) XP® Rabbit #46687. The virtual lane view (left) shows the target band (as indicated) and a band corresponding to Tau (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Simple Western™ analysis of lysates (1.0 mg/mL) from HEK 293 cells treated with UV (50 mJ, 30 min recovery) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation of phospho-tau (Thr217) protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb. Western blot analysis was performed using Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Western blot analysis of extracts from mouse brain, untreated (-) or λ phosphatase-treated (+), and rat brain using Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Simple Western™ analysis of lysates (1.0 mg/mL) from mouse brain using Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb #51625. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from control mIMCD-3 cells (lane 1) or p70 S6 kinase-1 knockout mIMCD-3 cells (lane 2), and control wild-type immortalized MEF cells (lane3) or p70 S6 Kinase-1/p70 S6 kinase-2 double knockout MEF cells (lane 4), using p70 S6 Kinase (E8K6T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the two knockout models confirms specificity of the antibody for p70 S6 kinase-1. Knockout cells were kindly provided by Dr. Mario Pende, INSERM, Paris, France.
Western blot analysis of extracts from various cell lines using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation of p70 S6 kinase protein from HeLa extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Western blot analysis was performed using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Western blot analysis of extracts from COS and 293 cells, untreated or UV-treated, using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (upper) or p38 MAPK Antibody #9212 (lower).
Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Tau (D1M9X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
Western blot analysis of extracts from 293 cells, untreated or UV-treated, NIH/3T3 cells, untreated or UV-treated and C6 cells, untreated or anisomycin-treated, using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.
Western blot analysis of extracts from normal mouse brain and Tau KO (-/-) mouse brain tissue using Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau KO (-/-) mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of normal mouse brain and Tau KO (-/-) mouse brain with Phospho-Tau (Ser214) (D1Q2X) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Tau-KO mouse brain tissue was kindly provided by Dr. Dominic Walsh at Brigham and Women's Hospital and Harvard Medical School.
Western blot analysis of extracts from various cell lines using p38 MAPK (D13E1) XP® Rabbit mAb.
Western blot analysis of extracts from serum-starved MCF7, NIH/3T3, and C6 cells, untreated (-) or treated with human IGF-1 (100 ng/ml, 10 min; +), using Phospho-p70 S6 Kinase (Thr389) (D5U1O) Rabbit mAb #97596 (upper), or p70 S6 Kinase (49D7) rabbit mAb #2708 (lower).
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines and tissues using Tau (D1M9X) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb in the presence of control peptide (left) or Phospho-SAPK/JNK (Thr183/Tyr185) Blocking Peptide #1215 (right).
Western blot analysis of extracts from mouse brain, untreated (-) or phosphatase-treated (+), using Phospho-Tau (Ser214) (D1Q2X) Rabbit mAb (upper) and Tau (D1M9X) Rabbit mAb #46687 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p38 MAPK (D13E1) XP® Rabbit mAb.
Western blot analysis of extracts from serum-starved 293T cells, untreated (-) or treated (+) with combinations of the following treatments as indicated: 20% fetal bovine serum (FBS) and λ phosphatase/calf intestinal alkaline phosphatase (CIP), using Phospho-p70 S6 Kinase (Thr389) (D5U1O) Rabbit mAb (upper) or p70 S6 Kinase Antibody #9202 (lower).
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human Alzheimer's brain using Tau (D1M9X) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded 293T cells untreated (left) or UV-treated (right) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using p38 MAPK (D13E1) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human pancreas using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, untreated (left) or UV-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human normal appendix using Tau (D1M9X) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using p38 MAPK (D13E1) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded normal human parathyroid using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded T-47D cell pellet (left, positive) or MDA-MB-231 cell pellet (right, negative) using Tau (D1M9X) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse testis using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Confocal immunofluorescent analysis of COS cells, untreated (left) or anisomycin-treated (right) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Immunohistochemical analysis of paraffin-embedded mouse lung using Tau (D1M9X) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with UV (100 mJ/cm2 with 30 min recovery; right), using p38 MAPK (D13E1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Immunohistochemical analysis of paraffin-embedded mouse kidney using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Anisomycin (25µM, 30 min; green) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (solid lines) or 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.
Confocal immunofluorescent analysis of fixed frozen mouse striatum using Tau (D1M9X) XP® Rabbit mAb (green), TMEM119 (E4B9S) Mouse mAb #98778 (red) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow cytometric analysis of HeLa cells using p38 MAPK (D13E1) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded mouse thymus using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Confocal immunofluorescent analysis of T-47D (positive, left) or MDA-MB-231 (negative, right) cells using Tau (D1M9X) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded mouse stomach using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse spleen using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using p70 S6 Kinase (E8K6T) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma (left), colon carcinoma (middle), or normal breast (right) using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (top) or p70 S6 Kinase Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on p70 S6 kinase protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded human small cell carcinoma of the salivary gland using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemical analysis of paraffin-embedded MEF cell pellet, wild-type (left, positive) or p70 S6 Kinase-1/ p70 S6 Kinase-2 double knockout (right, negative) using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Cell lines courtesy of Dr. Mario Pende, INSERM, Paris, France.
To Purchase # 89892
Cat. # Size Qty. Price
89892T
1 Kit  (8 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Tau (D1M9X) XP® Rabbit mAb 46687 20 µl
  • WB
  • IHC
  • IF
H M R 50-80 Rabbit IgG
Phospho-Tau (Ser214) (D1Q2X) Rabbit mAb 77348 20 µl
  • WB
  • IP
H M R 50-80 Rabbit IgG
Phospho-Tau (Thr217) (E9Y4S) Rabbit mAb 51625 20 µl
  • WB
  • IP
H M R 50-80 Rabbit IgG
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 4511 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk Mi Pg Sc 43 Rabbit IgG
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb 4668 20 µl
  • WB
  • IP
  • IHC
H M R Dm Sc 46, 54 Rabbit IgG
Phospho-p70 S6 Kinase (Thr389) (D5U1O) Rabbit mAb 97596 20 µl
  • WB
H M R Mk 70, 85 Rabbit IgG
p38 MAPK (D13E1) XP® Rabbit mAb 8690 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Hm Mk B Pg 40 Rabbit IgG
p70 S6 Kinase (E8K6T) XP® Rabbit mAb 34475 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R Mk 70, 85 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Phospho-Tau (Ser214/T217) Signaling Antibody Sampler Kit provides an economical means of detecting the signaling cascade leading to tau phosphorylation at Ser214 and Thr217, using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

Tau is a heterogeneous microtubule-associated protein that promotes and stabilizes microtubule assembly, especially in axons. Six isoforms with different amino-terminal inserts and different numbers of tandem repeats near the carboxy terminus have been identified, and tau is hyperphosphorylated at approximately 25 sites by Erk, glycogen synthase kinase-3 (GSK-3), and CDK5 (1,2). Phosphorylation decreases the ability of tau to bind to microtubules. Neurofibrillary tangles are a major hallmark of Alzheimer's disease (AD); these tangles are bundles of paired helical filaments (PHFs) composed of hyperphosphorylated tau. Furthermore, research studies have shown that inclusions of tau are found in a number of other neurodegenerative diseases, collectively known as tauopathies (1,3).

Numerous kinases, including p38 MAPK, JNK, CDK5, GSK3β, and SGK1, have been shown to phosphorylate tau at Ser214, which is found in AD and dementia with Lewy bodies (4-9). p38 MAPK participates in a signaling cascade controlling cellular responses to cytokines and stress and is activated by phosphorylation at Thr180 and Tyr182, allowing for downstream tau Ser214 phosphorylation (10-13). Another kinase that phosphorylates the Ser214 site on tau is the stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK). SAPK/JNK is preferentially activated by a variety of environmental stresses, including UV and gamma radiation, ceramides, inflammatory cytokines, and, in some instances, growth factors and GPCR agonists, which lead to phosphorylation of Thr183 and Tyr185 (14-19).

Tau phosphorylation at Thr217 has been identified as a highly specific biomarker of AD. Phosphorylated tau at Thr217 can be detected in cerebrospinal fluid in both preclinical and advanced stages of AD and has been shown to successfully differentiate between AD and non-AD neurodegenerative diseases (20,21). One kinase identified to phosphorylate the Thr217 site on tau is p70 S6 kinase (S6K1). S6K1 is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (22,23). Phosphorylation of Thr389 most closely correlates with S6K1 activity in vivo (24).

  1. Johnson, G.V. and Stoothoff, W.H. (2004) J Cell Sci 117, 5721-9.
  2. Hanger, D.P. et al. (1998) J Neurochem 71, 2465-76.
  3. Bramblett, G.T. et al. (1993) Neuron 10, 1089-99.
  4. Illenberger, S. et al. (1998) Mol Biol Cell 9, 1495-512.
  5. Götz, J. et al. (2001) Science 293, 1491-5.
  6. Yang, Y.C. et al. (2006) Mol Cell Biol 26, 8357-70.
  7. Liu, F. et al. (2006) FEBS Lett 580, 6269-74.
  8. Zhu, B. et al. (2010) Am J Physiol Lung Cell Mol Physiol 299, L493-501.
  9. Duka, V. et al. (2013) PLoS One 8, e75025.
  10. Rouse, J. et al. (1994) Cell 78, 1027-37.
  11. Han, J. et al. (1994) Science 265, 808-11.
  12. Lee, J.C. et al. Nature 372, 739-46.
  13. Freshney, N.W. et al. (1994) Cell 78, 1039-49.
  14. Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
  15. Ichijo, H. (1999) Oncogene 18, 6087-93.
  16. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
  17. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
  18. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  19. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.
  20. Barthélemy, N.R. et al. (2020) Alzheimers Res Ther 12, 26.
  21. Janelidze, S. et al. (2020) Nat Commun 11, 1683.
  22. Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82.
  23. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
  24. Weng, Q.P. et al. (1998) J Biol Chem 273, 16621-9.

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