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12594
HSP27 Antibody Sampler Kit
Primary Antibodies

HSP27 Antibody Sampler Kit #12594

Western Blotting Image 1

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.

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Western Blotting Image 2

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

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Western Blotting Image 3

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).

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Western Blotting Image 4

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).

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Western Blotting Image 5

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.

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IHC-P (paraffin) Image 6

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

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Western Blotting Image 7

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.

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IHC-P (paraffin) Image 8

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

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IHC-P (paraffin) Image 9

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

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IHC-P (paraffin) Image 10

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

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IHC-P (paraffin) Image 11

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

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IHC-P (paraffin) Image 12

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

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IHC-P (paraffin) Image 13

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).

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Flow Cytometry Image 14

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).

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IHC-P (paraffin) Image 15

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).

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IF-IC Image 16

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

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IHC-P (paraffin) Image 17

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

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Flow Cytometry Image 18

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

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IF-IC Image 19

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).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-HSP27 (Ser82) (D1H2F6) XP® Rabbit mAb 9709 20 µl
  • WB
  • IHC
  • IF
  • F
H M 27 Rabbit IgG
HSP27 (G31) Mouse mAb 2402 20 µl
  • WB
  • IHC
  • IF
H Mk 27 Mouse IgG1
Phospho-HSP27 (Ser15) Antibody 2404 20 µl
  • WB
H Mk 27 Rabbit 
Phospho-HSP27 (Ser78) Antibody 2405 20 µl
  • WB
  • IHC
  • F
H Mk 27 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

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.

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.

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.

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.
Entrez-Gene Id
3315
Swiss-Prot Acc.
P04792
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.

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