Product Name

Hsp70

Catalog #

SMC-100B

Alternative Names

Hsp70 1, Hsp70 2, Hsp70.1, Hsp72, HSPA1, HSPA1A, HSPA1B

Clone Number

C92F3A-5

Immunogen

Human Hsp70

Accession Number

NP_005336.3

Gene ID

3303

SwissProt

P08107

Applications

WB, IP, ELISA, ICC, IHC, FACS, IEM

Host Species

Mouse

Isotype

IgG1

Species Reactivity

Human, Mouse, Rat, Bovine, C.elegans, Canine, Chicken, Drosophilia, Carp, Guinea pig, Hamster, Monkey, Pig, Rabbit, Sheep

Recommended Dilutions

1μg/ml was sufficient for detection of Hsp70 in 20µof Hela cell lysate

Form

Protein G Purified

Storage Buffer

PBS pH7.2, 50% glycerol

Concentration

1mg/mL

Background Info

Detects a ~70kDa protein corresponding to the molecular mass of inducible Hsp70 on SDS PAGE immunoblots. The mapped epitope is in the region of amino acid residues 436-503. Does not cross-react with Hsc70 (Hsp73).

Conjugate

N/A

Package Size

200ug

Storage Temp

-20°C

Shipping Temp

Blue Ice or 4°C

Datasheet

SMC 100 Heat Shock Protein 70 (Hsp70)

Research Area

Chaperones, Heat Shock, Trafficking

Certificate of Analysis

1 μg/mL of SMC-100 was sufficient for detection of Hsp70 in 20μg of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Price

$309.00 USD Add to Cart

Mouse melanoma cells heat shocked and probed with C92 (SMC-100 was used at 1:1000 dilution in 1% BSA for a final conc. of 10ug/mL). 
Courtesy of Sklodowska, Curie Memorial Cancer Center and Institute of Oncology, Poland.


Western blot analysis of Hsp70 in cell lysates from 12 human cancer cell lines at 1:1000 dilution of SMC-100.


Human colon tissue stained (brown) in inflammatory cells.  Shown at a 1:100,000 dilution.

Research Background

Hsp70 genes encode abundant heat-inducible 70-kDa hsps (hsp70s). In most eukaryotes hsp70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 5O% identity (2). The N-terminal two thirds of hsp70s are more conserved than the C-terminal third. Hsp70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides (3). When hsc70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44 kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half (4). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (5). All hsp70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the hsp70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein (6). The universal ability of hsp70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport.

References

1. Welch W.J. and Suhan J.P. (1986) J Cell Biol. 103: 2035-2050.
2. Boorstein W. R., Ziegelhoffer T. & Craig E. A. (1993) J. Mol. Evol. 38(1): 1-17.
3. Rothman J. (1989) Cell 59: 591-601.
4. DeLuca-Flaherty et al. (1990) Cell 62: 875-887.
5. Bork P., Sander C. & Valencia A. (1992) Proc. Nut1 Acad. Sci. USA 89: 7290-7294.
6. Fink A.L. (1999) Physiol. Rev. 79: 425-449.
7. Galan A., et al. (2000) J. Biol. Chem. 275: 11418-11424.
8. Kondo T., et al. (2000) J. Biol. Chem. 275: 8872-8879.
9. Misaki T., et al. (1994) Clin. Exp. Immun. 98: 234-239.
10. Pockley A.G., et al. (1998) Immunol. Invest. 27: 367-377.
11. Moon I.S., et al. (2001) Cereb Cortex 11(3): 238-248.
12. Dressel et al. (2000) J. Immunol. 164: 2362-2371.
13. Verma A.K., et al. (2007) Fish and Shellfish Immunology. 22(5): 547-555.
14. Banduseela V.C., et al. (2009) Physiol Genomics. 39(3): 141-159.

Cited References

1. Anu Olkku, Jarkko J. Leskinen, Mikko J. Lammi, Kullervo Hynynen Anitta Mahonen. Ultrasound-induced activation of Wnt signaling in human MG-63 osteoblastic cells. Bone 47 (2010) 320–330.

2. Patricia Fernández-Llama, Sookkasem Khositseth, Patricia A Gonzales, Robert A Star, Trairak Pisitkun and Mark A Knepper. Tamm-Horsfall protein and urinary exosome isolation. Kidney International 77, 736-742 (April (2) 2010).

3. Ya-ming Xu, Marilyn T. Marron, Emily Seddon, Steven P. McLaughlin, Dennis T. Ray,Luke Whitesell and A. A. Leslie Gunatilaka. 2,3-Dihydrowithaferin A-3b-O-sulfate, a new potential prodrug of withaferin A from aeroponically grown Withania somnifera. Bioorganic & Medicinal Chemistry 17 (2009) 2210–2214

4. Masami Mitsuhashi, Masaru Yamaguchi, Tadashi Kojima, Ryo Nakajima and Kazutaka Kasai.Effects of HSP70 on the compression force-induced TNF-α and RANKL expression in human periodontal ligament cells. Inflammation Research Volume 60, Number 2, 187-194, DOI: 10.1007/s00011-010-0253-x

5. Liliana Batista-Nascimento, Daniel W. Neef, Phillip C. C. Liu, Claudina Rodrigues-Pousada, Dennis J. Thiele. Deciphering Human Heat Shock Transcription Factor 1 Regulation via Post-Translational Modification in Yeast. PLoS ONE 6(1): e15976. doi:10.1371/journal.pone.0015976

6. Sachie Marubayashi, Priya Koppikar, Tony Taldone, Omar Abdel-Wahab, Nathan West, Neha Bhagwat, Eloisi Caldas-Lopes, Kenneth N. Ross, Mithat Gönen, Alex Gozman, James H. Ahn, Anna Rodina, Ouathek Ouerfelli, Guangbin Yang, Cyrus Hedvat, James E. Bradner, Gabriela Chiosis and Ross L. Levine. HSP90 is a therapeutic target in JAK2-dependent myeloproliferative neoplasms in mice and humans. Published in Volume 120, Issue 10 (October 1, 2010) J Clin Invest. 2010;120(10):3578–3593. doi:10.1172/JCI42442.

7. Ya-ming Xu, Marilyn T. Marron, Emily Seddon, Steven P. McLaughlin, Dennis T. Ray, Luke Whitesell and A.A. Leslie Gunatilaka. 2,3-Dihydrowithaferin A-3β-O-sulfate, a new potential prodrug of withaferin A from aeroponically grown Withania somnifera. Bioorganic & Medicinal Chemistry Volume 17, Issue 6, 15 March 2009, Pages 2210-2214 Special Issue: Natural Products in Medicinal Chemistry. doi:10.1016/j.bmc.2008.10.091.

8. Shipp, C, Watson, K. and Jones, G. L. Associations of HSP90 Client Proteins in Human Breast Cancer. Anticancer Research June 2011 vol. 31 no. 6 2095-2101.

9. Julien Ochala, Ann-Marie Gustafson, Monica llano Diez, Guillaume Renaud, Meishan Li, Sudhakar Aare, Rizwan Qaisar, Varuna C Banduseela, Yvette Hedström, Xiaorui Tang, Barry Dworkin, Charles G Ford, Sreekumaran Nair Nair, Sue Perera, Mathias Gautel and Lars Larsson. Preferential skeletal muscle myosin loss in response to mechanical silencing in a novel rat intensive care unit model: Underlying mechanisms. The Journal of Physiology . February 14, 2011, doi: 10.1113/jphysiol.2010.202044.

10. Gunatilaka, Leslie, Wijeratne, Ekanayake Mudiyanselage Kithsiri, Xu, Ya-ming, Whitesell, Luke, Lindquist, Susan L. WITHAFERIN A ANALOGS AND USES THEREOF United States Patent Application 20110230551.

11. Noni T. Larkins, Robyn M. Murphy, and Graham D. Lamb. Absolute amounts and diffusibility of HSP72, HSP25 and αB-crystallin in fast- and slow-twitch skeletal muscle fibers of rat. October 2011, doi: 10.​1152/​ajpcell.​00266.​2011 Am J Physiol Cell Physiol. jpcell.00266.201

12. Kristina R Patterson , Sarah M. Ward , Benjamin Combs , Kellen Voss , Nicholas M Kanaan , Gerardo A Morfini , Scott T. Brady , Truman Christopher Gamblin , and Lester I. Binder. Heat Shock Protein 70 Prevents Both Tau Aggregation and the Inhibitory Effects of Pre-existing Tau Aggregates on Fast Axonal Transport. Biochemistry, DOI: 10.1021/bi200914.

13. Sandro Santagata , Ya-ming Xu , Kithsiri Wijeratne , Renee Kontnik , Christine Rooney , Casey Perley , Hyoungtae Kwon , Jon Clardy , Santosh Kesari , Luke Whitesell , Susan Lindquist , and Leslie Gunatilaka. Using the Heat-Shock Response to Discover Compounds that Target the Malignant Phenotype by Disrupting Protein Homeostasis. ACS Chem. Biol., DOI: 10.1021/cb200353m

14. Ying Zhang, Young-Hoon Ahn, Ivor J. Benjamin, Tadashi Honda, Ronald J. Hicks, Vittorio Calabrese, Philip A. Cole, Albena T. Dinkova-Kostova. HSF1-Dependent Upregulation of Hsp70 by Sulfhydryl-Reactive Inducers of the KEAP1/NRF2/ARE Pathway. Chemistry & Biology. Volume 18, Issue 11, 23 November 2011, Pages 1355-1361. doi:10.1016/j.chembiol.2011.09.