The heat shock proteins (HSPs) comprise a group of highly conserved, abundantly expressed proteins with diverse functions, including the assembly and sequestering of multiprotein complexes, transportation of nascent poly-peptide chains across cellular membranes and regulation of protein folding. Heat shock proteins (also known as molecular chaperones) fall into six general families: HSP 90, HSP 70, HSP 60, the low molecular weight HSPs, the immunophilins and the HSP 110 family. The HSP 110 family (also known as the HSP 105 family) is composed of HSP 105, Apg-1 and Apg-2. HSP 105 is a testis-specific and HSP 90-related protein. Research indicates that HSP 105 is specifically localized in the germ cells and may translocate into the nucleus after heat shock. It is suggested that HSP 105 may contribute to the stabilization of p53 proteins in the cytoplasm of the germ cells, preventing the potential induction of apoptosis by p53.
Background References
1. Bian Y., et al. 2014. An enzyme assisted RP-RPLC approach for in-depth analysis of human liver phosphoproteome. J. Proteomics 96:253-262.
2. Zhou H., et al. 2013. Toward a comprehensive characterization of a human cancer cell phosphoproteome. J. Proteome Res. 12:260-271.
Sequence Similarity
Belongs to the heat shock protein 70 family.
Tissue Specificity
Highly expressed in testis. Present at lower levels in most brain regions, except cerebellum. Overexpressed in cancer cells.
Post-translational Modification
Phosphorylation on Ser-509 may be important for regulation of the HSPA8/HSC70 chaperone activity.
Western blot analysis of Hsp105 on different lysates with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/1,000 dilution.
Lane 1: MCF7 cell lysate Lane 2: HepG2 cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: C6 cell lysate Lane 5: Mouse brain tissue lysate Lane 6: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 97 kDa Observed band size: 105 kDa
Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-88) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of Hsp105 on different lysates with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Hsp105 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 97 kDa Observed band size: 105 kDa
Exposure time: 15 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-88) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HepG2 cells labeling Hsp105 with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of NIH/3T3 cells labeling Hsp105 with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Immunocytochemistry analysis of C6 cells labeling Hsp105 with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp105 antibody (ET1612-88) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Flow cytometric analysis of HepG2 cells labeling Hsp105.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-88, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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