Application Data
(Establishing the GeneSwitch paradigm.(A) Western immunoblot demonstrated that the gmr-GS-GAL4 line specifically drove Hsp104 expression in the presence of RU486 (gmr-GS ON), but there was no expression of Hsp104 in the absence of the drug (gmr-GS OFF) at d7. The level of expression by gmr-GS was lower than with the other eye-specific drivers gmr-GAL4 and 1×gr-GAL4. Tubulin served as a loading control. Quantification of Western immunoblots confirmed that gmr-GS expressed at a lower level than the other drivers, with the amount of Hsp104 expressed by gmr-GS reaching about 33% of that expressed by 1×gr-GAL4. Hsp104 levels were normalized to tubulin (n = 3 (mean ± SEM)). (B) Paraffin sections demonstrate that the retinal tissue loss associated with gmr-MJDtrQ78 is apparent at d0, and progresses through d7. In comparison, control flies (7 d) display no such loss of retinal integrity. For each example shown here, a 7,000 um2 rectangular selection (used for quantification in Fig. 10) of a retinal section from three independent animals is presented. Each region was converted to a black and white image to show the area covered by tissue and quantitated by ImageJ analysis (see Methods). For the analysis in Fig. 10, regions from 6 independent animals were used for quantitation; all experiments were repeated at least three times with similar results.)
Rabbit anti-Yeast HSP104 Polyclonal Antibody | anti-HSP104 antibody
HSP104 (yeast) polyclonal antibody
Avoid freeze/thaw cycles.
Application Data
(Establishing the GeneSwitch paradigm.(A) Western immunoblot demonstrated that the gmr-GS-GAL4 line specifically drove Hsp104 expression in the presence of RU486 (gmr-GS ON), but there was no expression of Hsp104 in the absence of the drug (gmr-GS OFF) at d7. The level of expression by gmr-GS was lower than with the other eye-specific drivers gmr-GAL4 and 1×gr-GAL4. Tubulin served as a loading control. Quantification of Western immunoblots confirmed that gmr-GS expressed at a lower level than the other drivers, with the amount of Hsp104 expressed by gmr-GS reaching about 33% of that expressed by 1×gr-GAL4. Hsp104 levels were normalized to tubulin (n = 3 (mean ± SEM)). (B) Paraffin sections demonstrate that the retinal tissue loss associated with gmr-MJDtrQ78 is apparent at d0, and progresses through d7. In comparison, control flies (7 d) display no such loss of retinal integrity. For each example shown here, a 7,000 um2 rectangular selection (used for quantification in Fig. 10) of a retinal section from three independent animals is presented. Each region was converted to a black and white image to show the area covered by tissue and quantitated by ImageJ analysis (see Methods). For the analysis in Fig. 10, regions from 6 independent animals were used for quantitation; all experiments were repeated at least three times with similar results.)
Application Data
(Establishing the GeneSwitch paradigm.(A) Western immunoblot demonstrated that the gmr-GS-GAL4 line specifically drove Hsp104 expression in the presence of RU486 (gmr-GS ON), but there was no expression of Hsp104 in the absence of the drug (gmr-GS OFF) at d7. The level of expression by gmr-GS was lower than with the other eye-specific drivers gmr-GAL4 and 1×gr-GAL4. Tubulin served as a loading control. Quantification of Western immunoblots confirmed that gmr-GS expressed at a lower level than the other drivers, with the amount of Hsp104 expressed by gmr-GS reaching about 33% of that expressed by 1×gr-GAL4. Hsp104 levels were normalized to tubulin (n = 3 (mean ± SEM)). (B) Paraffin sections demonstrate that the retinal tissue loss associated with gmr-MJDtrQ78 is apparent at d0, and progresses through d7. In comparison, control flies (7 d) display no such loss of retinal integrity. For each example shown here, a 7,000 um2 rectangular selection (used for quantification in Fig. 10) of a retinal section from three independent animals is presented. Each region was converted to a black and white image to show the area covered by tissue and quantitated by ImageJ analysis (see Methods). For the analysis in Fig. 10, regions from 6 independent animals were used for quantitation; all experiments were repeated at least three times with similar results.)
Application Data
(Hsp104 delays aggregation of truncated MJD, but enhances aggregation of full-length MJD.(A and B) With the rh1-GAL4 driver at indicated time points, cryosections and IHC demonstrate accumulations of MJDtrQ78 and MJDnQ78 (red) over time, using anti-HA and anti-myc antibodies, respectively. Sections were co-stained with anti-Hsp104 or anti-Hsp70 (green) as indicated, and nuclei were labeled by Hoechst (blue). Hsp104 delayed but did not suppress aggregation of MJDtrQ78, but Hsp104 enhanced accumulation formation of MJDnQ78. Hsp70 suppressed aggregation of both MJD proteins. The size of the aggregates was quantified using ImageJ, with delineations for large inclusions (>5 um across), medium inclusions (2.5–5 um), or small inclusions ()
Application Data
(Hsp104 delays aggregation of truncated MJD, but enhances aggregation of full-length MJD.(A and B) With the rh1-GAL4 driver at indicated time points, cryosections and IHC demonstrate accumulations of MJDtrQ78 and MJDnQ78 (red) over time, using anti-HA and anti-myc antibodies, respectively. Sections were co-stained with anti-Hsp104 or anti-Hsp70 (green) as indicated, and nuclei were labeled by Hoechst (blue). Hsp104 delayed but did not suppress aggregation of MJDtrQ78, but Hsp104 enhanced accumulation formation of MJDnQ78. Hsp70 suppressed aggregation of both MJD proteins. The size of the aggregates was quantified using ImageJ, with delineations for large inclusions (>5 um across), medium inclusions (2.5–5 um), or small inclusions ()
Application Data
(ATPase activity and substrate binding are required for Hsp104 to modulate disease.(A) With the gmr-GAL4 driver at d7, expression of the inactive mutant UAS-Hsp104DPLDWB, which is unable to bind substrate or hydrolyze ATP, caused no effect on its own when expressed by gmr-GAL4. Additionally, the inactive Hsp104DPLDWB did not modulate the toxicity of MJDtrQ78 or MJDnQ78. Eye images and retinal sections showed moderate degeneration upon expression of MJDtrQ78 or MJDnQ78, but unlike wild-type Hsp104 (see Fig. 2), Hsp104DPDLWB did not mitigate the degeneration caused by MJDtrQ78 nor did it enhance the toxicity of MJDnQ78. Arrows indicate the width of the retina to highlight changes in tissue integrity. (B) Western immunoblot demonstrated that WT Hsp104 and Hsp104DPLDWB had similar expression levels. Tubulin served as a loading control. Quantification of Western immunoblots confirmed that protein expression levels were similar, but that Hsp104DPLDWB was expressed at levels slightly higher than WT Hsp104. Hsp104 signal was normalized to tubulin (n?=?3 (mean ± SEM)).)
Application Data
(ATPase activity and substrate binding are required for Hsp104 to modulate disease.(A) With the gmr-GAL4 driver at d7, expression of the inactive mutant UAS-Hsp104DPLDWB, which is unable to bind substrate or hydrolyze ATP, caused no effect on its own when expressed by gmr-GAL4. Additionally, the inactive Hsp104DPLDWB did not modulate the toxicity of MJDtrQ78 or MJDnQ78. Eye images and retinal sections showed moderate degeneration upon expression of MJDtrQ78 or MJDnQ78, but unlike wild-type Hsp104 (see Fig. 2), Hsp104DPDLWB did not mitigate the degeneration caused by MJDtrQ78 nor did it enhance the toxicity of MJDnQ78. Arrows indicate the width of the retina to highlight changes in tissue integrity. (B) Western immunoblot demonstrated that WT Hsp104 and Hsp104DPLDWB had similar expression levels. Tubulin served as a loading control. Quantification of Western immunoblots confirmed that protein expression levels were similar, but that Hsp104DPLDWB was expressed at levels slightly higher than WT Hsp104. Hsp104 signal was normalized to tubulin (n?=?3 (mean ± SEM)).)
Application Data
(Hsp104 delays aggregation of truncated MJD, but enhances aggregation of full-length MJD.(A and B) With the rh1-GAL4 driver at indicated time points, cryosections and IHC demonstrate accumulations of MJDtrQ78 and MJDnQ78 (red) over time, using anti-HA and anti-myc antibodies, respectively. Sections were co-stained with anti-Hsp104 or anti-Hsp70 (green) as indicated, and nuclei were labeled by Hoechst (blue). Hsp104 delayed but did not suppress aggregation of MJDtrQ78, but Hsp104 enhanced accumulation formation of MJDnQ78. Hsp70 suppressed aggregation of both MJD proteins. The size of the aggregates was quantified using ImageJ, with delineations for large inclusions (>5 um across), medium inclusions (2.5–5 um), or small inclusions ()
Application Data
(Hsp104 delays aggregation of truncated MJD, but enhances aggregation of full-length MJD.(A and B) With the rh1-GAL4 driver at indicated time points, cryosections and IHC demonstrate accumulations of MJDtrQ78 and MJDnQ78 (red) over time, using anti-HA and anti-myc antibodies, respectively. Sections were co-stained with anti-Hsp104 or anti-Hsp70 (green) as indicated, and nuclei were labeled by Hoechst (blue). Hsp104 delayed but did not suppress aggregation of MJDtrQ78, but Hsp104 enhanced accumulation formation of MJDnQ78. Hsp70 suppressed aggregation of both MJD proteins. The size of the aggregates was quantified using ImageJ, with delineations for large inclusions (>5 um across), medium inclusions (2.5–5 um), or small inclusions ()
Application Data
(Tuning Hsp104 expression level for the fly eye.(A) Expression of UAS-Hsp104 using gmr-GAL4 caused disruption of internal retinal structure by d7. A less strong driver, 1xgr-GAL4 minimized this effect and prevented disruption to cellular organization within the retina. Arrows indicate the width of the retina to highlight changes in tissue integrity. (B) Immunoblots demonstrated that 1×gr-GAL4 drives lower Hsp104 expression than gmr-GAL4 at d7. Actin served as a loading control. Quantitation of immunoblots determined that 1xgr-GAL4 levels of Hsp104 were 35% that of gmr-GAL4. Hsp104 levels were normalized to actin (n = 3 (mean +- SEM)).)
Application Data
(Tuning Hsp104 expression level for the fly eye.(A) Expression of UAS-Hsp104 using gmr-GAL4 caused disruption of internal retinal structure by d7. A less strong driver, 1xgr-GAL4 minimized this effect and prevented disruption to cellular organization within the retina. Arrows indicate the width of the retina to highlight changes in tissue integrity. (B) Immunoblots demonstrated that 1×gr-GAL4 drives lower Hsp104 expression than gmr-GAL4 at d7. Actin served as a loading control. Quantitation of immunoblots determined that 1xgr-GAL4 levels of Hsp104 were 35% that of gmr-GAL4. Hsp104 levels were normalized to actin (n = 3 (mean +- SEM)).)
WB (Western Blot)
(Western Blot Analysis of HSP104 (yeast), pAb (Prod. No. ADI-SPA-1040): Lane 1: MW Marker, Lane 2: Yeast Cell Lysate)
WB (Western Blot)
(Western Blot Analysis of HSP104 (yeast), pAb (Prod. No. ADI-SPA-1040): Lane 1: MW Marker, Lane 2: Yeast Cell Lysate)
WB (Western Blot)
(Western Blot Analysis of HSP104 (yeast), pAb: Lane 1: MW Marker, Lane 2: Yeast Cell Lysate)
WB (Western Blot)
(Western Blot Analysis of HSP104 (yeast), pAb: Lane 1: MW Marker, Lane 2: Yeast Cell Lysate)
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Product Notes
The HSP104 hsp104 (Catalog #AAA77020) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The HSP104 (yeast) polyclonal antibody reacts with Yeast and may cross-react with other species as described in the data sheet. AAA Biotech's HSP104 can be used in a range of immunoassay formats including, but not limited to, WB (Western Blot). Researchers should empirically determine the suitability of the HSP104 hsp104 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "HSP104, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
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