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9965
HSP/Chaperone Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

HSP/Chaperone Antibody Sampler Kit #9965

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HSP/Chaperone Antibody Sampler Kit: Image 1

Flow cytometric analysis of HeLa cells using HSP60 (D6F1) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

HSP/Chaperone Antibody Sampler Kit: Image 2

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 3

HeLa cells were either untreated (left panel) or heat shocked (right panel) for 1h. Chromatin immunoprecipitations were performed with cross-linked chromatin from cells and either HSF1 Antibody or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HSPA6 Promoter Primers #5551, human HSP70 intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

HSP/Chaperone Antibody Sampler Kit: Image 4

Flow cytometric analysis of A204 cells using BiP (C50B12) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

HSP/Chaperone Antibody Sampler Kit: Image 5

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HSP40 (C64B4) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 6

Flow cytometric analysis of HeLa cells using HSP90 (C45G5) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

HSP/Chaperone Antibody Sampler Kit: Image 7

Confocal immunofluorescent analysis of HeLa cells using Calnexin (C5C9) Rabbit mAb (green). Actin filaments have been labeled using DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

HSP/Chaperone Antibody Sampler Kit: Image 8

Confocal immunofluorescent analysis of NIH/3T3 cells using PDI (C81H6) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

HSP/Chaperone Antibody Sampler Kit: Image 9

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.

HSP/Chaperone Antibody Sampler Kit: Image 10

Confocal immunofluorescent analysis of A-204 cells using HSP60 (D6F1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

HSP/Chaperone Antibody Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 12

Flow cytometric analysis of K562 cells, using HSF1 Antibody (blue) compared to a nonspecific negative control antibody (red).

HSP/Chaperone Antibody Sampler Kit: Image 13

Immunohistochemical analysis of paraffin-embedded human glioblastoma using BiP (C50B12) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 14

Western blot analysis of extracts from HeLa, C2C12, C6 and COS cells using HSP40 (C64B4) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 15

Confocal immunofluorescent analysis of HeLa cells using HSP90 (C45G5) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).

HSP/Chaperone Antibody Sampler Kit: Image 16

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Calnexin (C5C9) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

HSP/Chaperone Antibody Sampler Kit: Image 17

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using PDI (C81H6) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded mouse lung using HSP60 (D6F1) XP® Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing cytoplasmic localization using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 20

DAPI staining (left) and immunofluorescent staining (right) of paraformaldehyde-fixed HeLa cells, using HSF1 antibody.

HSP/Chaperone Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using BiP (C50B12) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 22

Immunohistochemical analysis of paraffin-embedded human colon using HSP90 (C45G5) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 23

Western blot analysis of extracts from PANC1, HepG2 and A204 cells using Calnexin (C5C9) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 24

Immunohistochemical analysis of paraffin-embedded human lymphoma using PDI (C81H6) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 25

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HSP60 (D6F1) XP® Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 26

Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 27

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HSF1 Antibody in the presence of control peptide (left) or antigen-specific peptide (right).

HSP/Chaperone Antibody Sampler Kit: Image 28

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using BiP (C50B12) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 29

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HSP90 (C45G5) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 30

Immunohistochemical analysis of paraffin-embedded mouse spleen using PDI (C81H6) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 31

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using HSP60 (D6F1) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

HSP/Chaperone Antibody Sampler Kit: Image 32

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 33

Immunohistochemical analysis of paraffin-embedded human pituitary adenoma, using HSF1 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 34

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using BiP (C50B12) Rabbit mAb in the presence of control peptide (left) or BiP Blocking Peptide #1084 (right).

HSP/Chaperone Antibody Sampler Kit: Image 35

Western blot analysis of extracts from HeLa, NIH/3T3, PC12 and COS cells using HSP90 (C45G5) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 36

Western blot analysis of extracts from various cell types using PDI (C81H6) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 37

Western blot analysis of extracts from various cell lines using HSP60 (D6F1) XP® Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 38

Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using HSP70 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 39

Immunohistochemical analysis of paraffin-embedded human breast carcinoma showing nuclear localization, using HSF1 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 40

Western blot analysis of extracts from various cell lines using BiP (C50B12) Rabbit mAb.

HSP/Chaperone Antibody Sampler Kit: Image 41

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using HSF1 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 42

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using HSF1 Antibody.

HSP/Chaperone Antibody Sampler Kit: Image 43

Western blot analysis of extracts from HeLa, NIH/3T3, C6 and COS cells, using HSF1 antibody.

To Purchase # 9965T
Product # Size Price
9965T
1 Kit  (8 x 20 µl) $ 554

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
HSP60 (D6F1) XP® Rabbit mAb 12165 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Hm Mk X Z B Pg 60 Rabbit IgG
HSP70 Antibody 4872 20 µl
  • WB
  • IHC
H M R Mk B 72, 73 Rabbit 
HSF1 Antibody 4356 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 82 Rabbit 
BiP (C50B12) Rabbit mAb 3177 20 µl
  • WB
  • IHC
  • F
H M 78 Rabbit IgG
HSP40 (C64B4) Rabbit mAb 4871 20 µl
  • WB
  • IHC
H M R Mk 40 Rabbit 
HSP90 (C45G5) Rabbit mAb 4877 20 µl
  • WB
  • IHC
  • IF
  • F
H M R Mk 90 Rabbit IgG
Calnexin (C5C9) Rabbit mAb 2679 20 µl
  • WB
  • IHC
  • IF
H Mk 90 Rabbit IgG
PDI (C81H6) Rabbit mAb 3501 20 µl
  • WB
  • IHC
  • IF
H M R Mk 57 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The HSP/Chaperone Sampler Kit provides an economical means to investigate protein folding within the cell. The kit contains enough primary and secondary antibodies to perform two Western blot experiments with each antibody.

Specificity / Sensitivity

HSP40 (C64B4) Rabbit mAb detects endogenous levels of total HSP40 protein. HSP60 (D6F1) XP® Rabbit mAb recognizes endogenous levels of total HSP60 protein. HSP70 Antibody detects endogenous levels of total HSP70 protein (HSP70-Hom, HSP70-1). HSP90 (C45G5) Rabbit mAb detects endogenous levels of total HSP90 protein. HSF1 Antibody detects endogenous levels of total HSF1 protein. Calnexin (C5C9) Rabbit mAb detects endogenous levels of total calnexin protein. PDI (C81H6) Rabbit mAb detects endogenous levels of total PDI protein. BiP (C50B12) Rabbit mAb detects endogenous levels of total BiP protein. Each of these antibodies recognizes only its specific target.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to human HSP70, and corresponding to residues at the carboxy-terminus of human HSF1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Rabbit monclonal antibodies are produced by immunizing rabbits with a synthetic peptide corresponding to residues surrounding Trp68 of human HSP60 protein, residues surrounding Gly584 of human BiP, surrounding Asn300 of HSP90, corresponding to Glu223 of human HSP40/Hdj1, corresponding to the sequence of human calnexin, and corresponding to the sequence of human PDI.

Background

HSP70 and HSP90 are molecular chaperones expressed constitutively under normal conditions to maintain protein homeostasis and are induced upon environmental stress (1). HSP70 and HSP90 interact with unfolded proteins to prevent irreversible aggregation and catalyze the refolding of their substrates in an ATP-dependent manner (1). HSP40 family proteins bind unfolded proteins and prevent their aggregation, and deliver unfolded protiens to HSP70 (2). HSP60 has primarily been known as a mitochondrial protein that is important for folding key proteins after import into the mitochondria (3). HSP60 is also present in the cytosol of many cells and is induced by stress, inflammatory and immune responses, autoantibodies correlated with Alzheimer's, coronary artery diseases, MS, and diabetes (4-7). Secretory and transmembrane proteins are synthesized on polysomes and translocate into the endoplasmic reticulum (ER) where they are often modified by the formation of disulfide bonds, amino-linked glycosylation and folding. The ER contains a pool of molecular chaperones including calnexin, BiP and protein disulfide isomerase (PDI). Calenxin is a calcium-binding protein embedded in the ER membrane that retains newly synthesized glycoproteins inside the ER to ensure proper folding and quality control (8,9). When protein folding is disturbed inside the ER, Bip synthesis is increased. Subsequently, BiP binds to misfolded proteins to prevent them from forming aggregates and assists them to refold properly (10). PDI catalyzes the formation and isomerization of disulfide bonds required to reach a proteins native state (11). Heat shock gene transcription is regulated by a familly of heat shock factors (HSFs), transcriptional activators that bind to heat shock response elements (HSEs) located upstream of all heat shock genes (12). During attenuation from the heat shock response, HSF1 is repressed by direct binding of HSP70, HSP40/Hdj-1 and HSF binding protein 1 (HSBP1) (13).

  1. Nollen, E.A. and Morimoto, R.I. (2002) J. Cell Sci. 115, 2809-2816.
  2. Fan, C.Y. et al. (2003) Cell Stress Chaperones 8, 309-316.
  3. Jindal, S. et al. (1989) Mol. Cell Biol. 9, 2279-2283.
  4. Itoh, H. et al. (2002) Eur. J. Biochem. 269, 5931-5938.
  5. Gupta, S. and Knowlton, A.A. J. Cell Mol. Med. 9, 51-58.
  6. Deocaris, C.C. et al. (2006) Cell Stress Chaperones 11, 116-128.
  7. Lai, H.C. et al. (2007) Am. J. Physiol. Endocrinol. Metab. 292, E292-E297.
  8. Bergeron, J.J. et al. (1994) Trends Biochem. Sci. 19, 124-128.
  9. Williams, D.B. (2006) J. Cell Sci. 119, 615-623.
  10. Kohno, K. et al. (1993) Mol Cell Biol 13, 877-90.
  11. Ellgaard, L. and Ruddock, L.W. (2005) EMBO Rep. 6, 28-32.
  12. Morimoto, R.I. (1998) Genes Dev 12, 3788-96.
  13. Satyal, S.H. et al. (1998) Genes Dev 12, 1962-74.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.