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12594
HSP27 Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

HSP27 Antibody Sampler Kit #12594

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HSP27 Antibody Sampler Kit: Image 1

Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

HSP27 Antibody Sampler Kit: Image 2

Confocal immunofluorescent analysis of A549 cells using HSP27 (G31) Mouse mAb (green). Red = Propidium iodide (fluorescent DNA dye).

HSP27 Antibody Sampler Kit: Image 3

Western blot analysis of extracts from HeLa cells (lane 1) or HSP27 knock-out cells (lane 2) using HSP27 (G31) Mouse mAb #2402 (upper), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the HSP27 knock-out HeLa cells confirms specificity of the antibody for HSP27.

HSP27 Antibody Sampler Kit: Image 4

Western blot analysis of extracts from HeLa and COS cells, untreated, anisomycin-treated or UV-treated, using Phospho-HSP27 (Ser15) Antibody (upper) or HSP27 (G31) Monoclonal Antibody #2402 (lower).

HSP27 Antibody Sampler Kit: Image 5

Flow cytometric analysis of HeLa cells, untreated (blue) or UV treated (green), using Phospho-HSP27 (Ser78) Antibody compared to a nonspecific negative control antibody (red).

HSP27 Antibody Sampler Kit: Image 6

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.

HSP27 Antibody Sampler Kit: Image 7

Confocal immunofluorescent analysis of C2C12 cells, untreated (left) or treated with λ phosphatase (middle), and NIH/3T3 cells (right) using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Negative staining in NIH/3T3 cells is in agreement with the observation that NIH/3T3 cells do not express HSP27 under basal conditions (5,7).

HSP27 Antibody Sampler Kit: Image 8

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using HSP27 (G31) Mouse mAb.

HSP27 Antibody Sampler Kit: Image 9

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-HSP27 (Ser78) Antibody.

HSP27 Antibody Sampler Kit: Image 10

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

HSP27 Antibody Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left panels) or lambda phosphatase-treated (right panels), using Phospho-HSP27 (Ser82) Antibody #2401 (upper panels) or HSP27 (G31) Mouse mAb (lower panels).

HSP27 Antibody Sampler Kit: Image 12

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-HSP27 (Ser78) Antibody.

HSP27 Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

HSP27 Antibody Sampler Kit: Image 14

Western blot analysis of extracts from HeLa and COS cells, untreated, anisomycin-treated or UV-treated, using Phospho-HSP27 (Ser78) Antibody (upper) or HSP27 (G31) mAb #2402 (lower).

HSP27 Antibody Sampler Kit: Image 15

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or UV-treated (right), using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

HSP27 Antibody Sampler Kit: Image 16

Western blot analysis of extracts from HeLa and COS-7 cells using HSP27 (G31) Mouse mAb.

HSP27 Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

HSP27 Antibody Sampler Kit: Image 18

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® HSP27 siRNA I (+) using HSP27 (G31) Mouse mAb and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107. The HSP27 (G31) Mouse mAb confirms silencing of HSP27 expression, while the p44/42 MAPK (Erk1/2) (3A7) Mouse mAb is used to control for loading and specificity of HSP27 siRNA.

HSP27 Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

HSP27 Antibody Sampler Kit: Image 20

Western blot analysis of extracts from HeLa or HT-29 cells, untreated (-) or treated (+) with either UV (40 mJ/cm2 with 30 min recovery) or anisomycin (25 μg/mL, 30 min), using Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb.

To Purchase # 12594T
Product # Size Price
12594T
1 Kit  (4 x 20 µl) $ 327

Product Description

The HSP27 Antibody Kit provides an economical means to evaluate the activation status of the HSP27 protein. The kit contains enough primary antibody to perform two western blot experiments per primary antibody.

Specificity / Sensitivity

Each antibody in the HSP27 Antibody Sampler Kit recognizes endogenous levels of its specific target. Activation state antibodies detect their intended targets only when phosphorylated at the indicated site.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser82 of human HSP27 and to full-length human HSP27 protein. Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser15 or Ser78 of human HSP27 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. Like other small HSPs, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the HSP27 expression increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78, and Ser82 by MAPKAPK-2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6).

  1. Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
  2. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
  3. Rouse, J. et al. (1994) Cell 78, 1027-1037.
  4. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
  5. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
  6. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

Limited Uses

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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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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