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9965
HSP/Chaperone Antibody Sampler Kit
Primary Antibodies

HSP/Chaperone Antibody Sampler Kit #9965

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

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

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

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

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

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

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

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

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.

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

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

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

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

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

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

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).

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

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

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

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

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

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

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).

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

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).

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

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

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

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

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

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.

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

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

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).

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

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).

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).

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

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

Immunohistochemical analysis of frozen SKOV-3 xenograft using BiP (C50B12) Rabbit mAb.

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

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.

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

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.

To Purchase # 9965T
Product # Size Price
9965T
1 Kit  (8 x 20 µl) N/A

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 
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. Morimoto, R.I. (1998) Genes Dev 12, 3788-96.
  3. Fan, C.Y. et al. (2003) Cell Stress Chaperones 8, 309-316.
  4. Bergeron, J.J. et al. (1994) Trends Biochem Sci 19, 124-8.
  5. Jindal, S. et al. (1989) Mol Cell Biol 9, 2279-83.
  6. Williams, D.B. (2006) J Cell Sci 119, 615-23.
  7. Ellgaard, L. and Ruddock, L.W. (2005) EMBO Rep. 6, 28-32.
  8. Itoh, H. et al. (2002) Eur. J. Biochem. 269, 5931-5938.
  9. Gupta, S. and Knowlton, A.A. J. Cell Mol. Med. 9, 51-58.
  10. Deocaris, C.C. et al. (2006) Cell Stress Chaperones 11, 116-128.
  11. Satyal, S.H. et al. (1998) Genes Dev 12, 1962-74.
  12. Lai, H.C. et al. (2007) Am. J. Physiol. Endocrinol. Metab. 292, E292-E297.
  13. Kohno, K. et al. (1993) Mol Cell Biol 13, 877-90.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.

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