Hap70 antibody

Antigen: Hap70

Clonality: Polyclonal

Host: Rabbit

Reactivity: Chlamydomonas reinhardtii (C. reinhardtii)

Application: Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN361872

Additional Information

Immunogen: Mature Hsp70B protein expressed with N- and C- terminal hexahistidine tags in E.coli, purified with Ni-NTA

Format: Antiserum

Description: Hsp70B (heat shock protein 70B) is a nuclear-encoded, chloroplast-targeted chaperone of the Hsp70 family. It is the major Hsp70 in the stroma of Chlamydomonas chloroplasts. It interacts with Hsp90C, CGE1, CDJ2 and VIPP1 (1-3). It has been shown that using Chlamydomonas reinhardtii as plant model organism the alga encodes a HEP homolog (termed HEP2) that is localized to the stroma (4). HEP2 is expressed constitutively as a low abundance protein with an apparent molecular mass of ~21 kDa. In cell extracts HEP2 interacts with HSP70B in an ATP-dependent fashion. Coexpression of HSP70B with HEP2 in E. coli yields high levels of CGE1-binding competent HSP70B, which also displayed ATPase activity (4). Inactive HSP70B was more prone to proteolysis than active HSP70B. Although inactive HSP70B interacts with HEP2, it is not activated. Active HSP70B remains active for 48 h in the absence of HEP2, suggesting that HEP2 was not involved in maintaining HSP70B in an active state. However, it was found that some HSP70B expressed as a fusion protein with an N-terminal extension was activated when HEP2 was present during cleavage of the fusion protein, suggesting that in vivo HEP2 might be required for renewed folding of HSP70B after transit peptide cleavage (4).

Specificity: Detects ~70kDa. Species cross-reactivity: Chlamydomonas reinhardtii. Other species not yet tested.

Application Details

Application Notes: Recommended dilution for WB 1:10,000 with ECL

Buffer: Lyophilized rabbit serum. (For reconstitution add 100uL of sterile water).

Storage: -20°C, 1 year+, shipped on cold packs or ambient

Restrictions: For Research Use only

Publications

Publications: Schroda, Vallon, Whitelegge et al.: "The chloroplastic GrpE homolog of Chlamydomonas: two isoforms generated by differential splicing." in: The Plant cell, Vol. 13, Issue 12, pp. 2823-39, 2001 (PubMed).

Liu, Willmund, Whitelegge et al.: "J-domain protein CDJ2 and HSP70B are a plastidic chaperone pair that interacts with vesicle-inducing protein in plastids 1." in: Molecular biology of the cell, Vol. 16, Issue 3, pp. 1165-77, 2005 (PubMed).