Revision 1

#12651Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-PDGF Receptor β (Tyr751) (C63G6) Rabbit mAb 4549 20 µl 190 kDa Rabbit IgG
PDGF Receptor β (28E1) Rabbit mAb 3169 20 µl 190 kDa Rabbit IgG
Phospho-SHP-2 (Tyr542) Antibody 3751 20 µl 72 kDa Rabbit 
SHP-2 (D50F2) Rabbit mAb 3397 20 µl 72 kDa Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb 4060 20 µl 60 kDa Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb 4691 20 µl 60 kDa Rabbit IgG
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 4370 20 µl 44, 42 kDa Rabbit IgG
p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 4695 20 µl 42, 44 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The PDGF Receptor Activation Antibody Sampler Kit provides an economical means to evaluate the activation status of multiple members of the PDGF receptor pathway, including SHP-2, Akt, and p44/42 MAPK (Erk1/2). The kit includes enough antibody to perform two western blot experiments per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Platelet derived growth factor (PDGF) family proteins form dimers (PDGF AA, PDGF AB, PDGF BB, PDGF CC, and PDGF DD) that bind receptor tyrosine kinases PDGF receptor α (PDGFRα) and PDGF receptor β (PDGFRβ) in a specific pattern. PDGFRβ homodimers bind PDGF BB and DD homodimers and the PDGF AB heterodimer. Heteromeric receptor PDGF α/β binds PDGF B, C, and D homodimers and the PDGF AB heterodimer (1). Ligand binding induces PDGF receptor dimerization and autophosphorylation, followed by binding and activation of cytoplasmic SH2 domain-containing signal transduction molecules, such as GRB2, Src, GAP, PI3 kinase, PLCγ, and NCK. Activated PDGF receptors initiate signaling pathways that control cell growth, actin reorganization, migration, and differentiation (2). PDGFRβ kinase-insert region residue Tyr751 forms the PI3 kinase docking site, and phosphorylation of PDGFRβ at this site inhibits the association between the SH2 domain of the PI3 kinase p85 subunit and PDGFRβ (3,4).
SHP-2 (PTPN11) is a nonreceptor protein tyrosine phosphatase that participates in signaling pathways that control cell growth, differentiation, migration, and death (5). Activation of SHP-2 and its association with Gab1 is critical for sustained Erk activation downstream of growth factor receptors and cytokines (6). Phosphorylation of SHP-2 at Tyr542 and Tyr580 in response to growth factor receptor activation is thought to relieve basal inhibition and stimulate SHP-2 tyrosine phosphatase activity (7,8).
Insulin and various growth/survival factors activate Akt, a kinase that acts in a wortmannin-sensitive pathway involving PI3 kinase to help control survival and apoptosis (9-11). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (12) and by phosphorylation within the carboxy terminus at Ser473.
The p44/42 MAPK (Erk1/2) signaling pathway is activated in response to extracellular stimuli including mitogens, growth factors, and cytokines (13-15). Research suggests that this pathway is an important target in cancer diagnosis and treatment (16). External stimuli lead to activation of a kinase cascade that results in the activation of p44 and p42 by a MAP kinase. MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively.
Clinical studies describe PDGF expression in a number of different solid tumors, from glioblastomas to prostate carcinomas. The biological role of PDGF signaling in these tumors varies from autocrine stimulation of cancer cell growth to more subtle paracrine interactions involving adjacent stroma and even angiogenesis. Targeting PDGF signaling may be an effective way for tumor treatment (17).

  1. Deuel, T.F. et al. (1988) Biofactors 1, 213-7.
  2. Ostman, A. and Heldin, C.H. (2001) Adv Cancer Res 80, 1-38.
  3. Betsholtz, C. et al. (2001) Bioessays 23, 494-507.
  4. Ramalingam, K. et al. (1995) Bioorg Med Chem 3, 1263-72.
  5. Qu, C.K. (2000) Cell Res 10, 279-88.
  6. Maroun, C.R. et al. (2000) Mol Cell Biol 20, 8513-25.
  7. Bennett, A.M. et al. (1994) Proc Natl Acad Sci U S A 91, 7335-9.
  8. Lu, W. et al. (2001) Mol Cell 8, 759-69.
  9. Franke, T.F. et al. (1997) Cell 88, 435-7.
  10. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
  11. Franke, T.F. et al. (1995) Cell 81, 727-36.
  12. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
  13. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
  14. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
  15. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
  16. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
  17. George, D. (2001) Semin Oncol 28, 27-33.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #12651

    PDGF Receptor Activation Antibody Sampler Kit

    PDGF Receptor Activation Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from 3T3 cells using PDGF Receptor β (28E1) Rabbit mAb #3169. The virtual lane view (left) shows a single 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 2 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with PDGF Receptor β (28E1) Rabbit mAb (green, left), β3-Tubulin (E9F3E) Mouse mAb #45058 (red, right), and DAPI #4083 (blue, right).
    PDGF Receptor Activation Antibody Sampler Kit: Image 3 Expand Image
    Confocal immunofluorescent analysis of fixed frozen mouse cerebral cortex labeled with PDGF Receptor β (28E1) Rabbit mAb (green, left), β3-Tubulin (E9F3E) Mouse mAb #45058 (red, right), and DAPI #4083 (blue, right).
    PDGF Receptor Activation Antibody Sampler Kit: Image 4 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with Calyculin A (100 uM, 30 min) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060. The virtual lane view (left) shows a single 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 5 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved HeLa cells treated with TPA (400 nM, 4 hours) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370. The virtual lane view (left) shows the 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 6 Expand Image
    Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using Akt (pan) (C67E7) Rabbit mAb #4691
    PDGF Receptor Activation Antibody Sampler Kit: Image 8 Expand Image
    Immunoprecipitation of Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb. Western blot analysis was performed using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as a secondary antibody.
    PDGF Receptor Activation Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines, using PDGF Receptor β (28E1) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from control 293T cells (lane 1) or SHP-2 knockout 293T cells (lane 2) using SHP-2 (D50F2) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SHP-2 knockout 293T cells confirms specificity of the antibody for SHP-2.
    PDGF Receptor Activation Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from C6 and NIH/3T3 cells, starved for 18 hours and either untreated or PDGF-treated (50ng/ml, 20 minutes), using Phospho-SHP-2 (Tyr542) Antibody (upper) or control SHP-2 Antibody #3752 (lower).
    PDGF Receptor Activation Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
    PDGF Receptor Activation Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from COS cells, untreated or treated with either U0126 #9903 (10 µM for 1h) or TPA #4174 (200 nM for 10 m), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107.
    PDGF Receptor Activation Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts of NIH/3T3 cells, untreated or stimulated with Platelet-Derived Growth Factor (PDGF) #9909, using Phospho-PDGF Receptor-β (Tyr751) (C63G6) Rabbit mAb (upper) or PDGF Receptor-β (2B3) Mouse mAb #3175 (lower).
    PDGF Receptor Activation Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts from various cell lines using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 17 Expand Image
    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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 18 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PDGF Receptor β (28E1) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 19 Expand Image
    Western blot analysis of extracts from various cell lines using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of phospho-Akt (Ser473) from Jurkat extracts treated with Calyculin A #9902 (100nM, 30 min). Lane 1 is 10% input, lane 2 is Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was performed with Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    PDGF Receptor Activation Antibody Sampler Kit: Image 21 Expand Image
    Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA #4174 as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
    PDGF Receptor Activation Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 23 Expand Image
    Western blot analysis of extracts from Hek 293 cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p44/42 MAPK (Erk1/2) siRNA (+), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 and α-Tubulin (11H10) Rabbit mAb #2125. The p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb confirms silencing of p44/42 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p44/42 MAPK (Erk1/2) siRNA.
    PDGF Receptor Activation Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded human glioblastoma using PDGF Receptor β (28E1) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using SHP-2 (D50F2) Rabbit mAb (left) or SHP-2 Antibody (right). These two antibodies detect independent, unique epitopes on human SHP-2. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
    PDGF Receptor Activation Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 28 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (C67E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) Blocking Peptide #1085 (right).
    PDGF Receptor Activation Antibody Sampler Kit: Image 29 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic and nuclear localization, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 30 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-87MG cells, showing membrane localization, using PDGF Receptor β (28E1) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 33 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 34 Expand Image
    Immunohistochemical analysis using Akt (pan) (C67E7) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
    PDGF Receptor Activation Antibody Sampler Kit: Image 35 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded K-562 cell pellet (left, high-expressing) or Saos-2 cell pellet (right, low-expressing) using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)
    PDGF Receptor Activation Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb on SignalSlide Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103 (paraffin-embedded NIH/3T3 cells, treated with U0126 #9903 (left) or TPA #4174 (right).
    PDGF Receptor Activation Antibody Sampler Kit: Image 39 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Akt (pan) (C67E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
    PDGF Receptor Activation Antibody Sampler Kit: Image 40 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb in the presence of control peptide (left) or #1240 p44/42 MAPK (Erk1/2) Blocking Peptide (#4695 Specific) (right).
    PDGF Receptor Activation Antibody Sampler Kit: Image 41 Expand Image
    Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or PDGF-treated (right), using PDGF Receptor beta (28E1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PDGF Receptor Activation Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 43 Expand Image
    Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.
    PDGF Receptor Activation Antibody Sampler Kit: Image 44 Expand Image
    Confocal immunofluorescent analysis of Drosophila egg chambers, untreated (top) or λ phosphatase-treated (bottom), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PDGF Receptor Activation Antibody Sampler Kit: Image 45 Expand Image
    Confocal immunofluorescent analysis of NIH/3T3 cells, treated with either U0126 (MEK1/2 Inhibitor) #9903 (left) or PDGF (right), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    PDGF Receptor Activation Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded human melanoma using using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.
    PDGF Receptor Activation Antibody Sampler Kit: Image 48 Expand Image
    Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PDGF Receptor Activation Antibody Sampler Kit: Image 49 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse ovary using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 50 Expand Image
    Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 51 Expand Image
    Flow cytometric analysis of Jurkat cells, treated with U0126 (10 µM, 2 hrs; green) or treated with TPA #4174 (200 nM, 30 min; blue) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 52 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using SHP-2 (D50F2) Rabbit mAb.
    PDGF Receptor Activation Antibody Sampler Kit: Image 53 Expand Image
    Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
    PDGF Receptor Activation Antibody Sampler Kit: Image 54 Expand Image
    Flow cytometric analysis of Jurkat cells using p44/42 MAPK (Erk1/2) (137F5) 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 55 Expand Image
    Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
    PDGF Receptor Activation Antibody Sampler Kit: Image 56 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue) using Phospho-Akt (Ser473) (D9E) 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.
    PDGF Receptor Activation Antibody Sampler Kit: Image 57 Expand Image
    Immunoprecipitation of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) from 3T3 cell extracts. Cells were treated with TPA, (200 nM, 15 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb. Western blot was performed using Phosphop44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (D4W3E) mAb #45262 was used as a secondary antibody.