Application
Western blot of mouse HSP 84usingApplication
Immunofluorescent analysis of Heat Shock Protein 86 (HSP86, green) in HeLa cells and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a HSP86 polyclonal antibody , at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.Application
Immunohistochemistry was performed on differentiated colon adenocarcinoma from paraffin-embedded human colon tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA in PBST for 30 minutes at room temperature and then probed with a Heat Shock Protein 86 (Hsp86) polyclonal antibody at a dilution of 1:100 for 1 hour in a humidified chamber (right panel). As a negative control, the primary antibody was eliminated from the staining procedure (left panel). Tissues were washed extensively with PBS/0.025% Tween-20 and endogenous peroxidase activity quenched with Peroxidase Suppressor for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:250 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit. Tissues were counterstained with hematoxylin and prepped for mounting.Application
Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody at a dilution of 1:1000 overnight at 4ºC on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using ECL Substrate.Application
Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.Application
Western blot analysis of Heat Shock Protein 86 (HSP86) was performed by loading 50ug of the indicated whole cell lysates and 15ul of PageRuler Prestained Protein Ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSP86 polyclonal antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performedApplication
Immunohistochemistry was performed on normal deparaffinized Human kidney tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.Application
Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.Application
Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an AP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using Lumi-Phos Substrate.Application
Western blot analysis of epidermoid carcinoma (A431) cells using Heat Shock Protein 86 Polyclonal Antibody . Decreasing amounts of A431 whole cell lysates were probed with PA3-013 at a dilution of 1:2000 followed by an anti-rabbit IgG secondary antibody and SuperSignal West Pico Chemiluminescent Substrate. Lanes 1 and 2 are control samples of purified, recombinant Hsp86 (rHsp86) and Hsp84 (rHsp84), respectively. Data courtesy of the Innovators Program.| Product Name | Heat Shock Protein 86 antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit Polyclonal antibody to Heat Shock Protein 86Heat shock proteins (HSP) are expressed in response to various biological stresses, including heat. HSP90 is a 90 kDa protein that is induced under stress conditions, but is also one of the most abundant cellular proteins found under non-stress conditions. HSP90 has been found to be associated with a number of other intracellular proteins, including steroid receptors, actin, tubulin, Ah receptor, and some kinases. Studies have shown that murine HSP90 exists as two forms, HSP84 and HSP86, coded by related but separate genes, with 86% homologous amino acid sequences. These forms are analogous to the two forms of human HSP90, HSP89 beta and HSP89 alpha. In an unstressed mouse fibroblast, the basal level of HSP84 is found to be double that of HSP86. However, after heat shock, HSP86 shows a greater increase. Studies also suggest that upon cellular differentiation, the level of HSP86, but not HSP84, decreases. HSP84 and HSP86, which may be subject to estrogenic regulation, have been found as components of the non-DNA binding form of mouse glucocorticoid receptor, but dissociated from the transformed DNA-binding form. |
| Immunogen | Synthetic peptide corresponding to residues P(2) E E T Q T Q D Q P M(12) of mosue HSP86. |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCIHC-FIHC-PIPWB |
| IHC-P: 5 µg/ml: |
|
| Species Reactivity | HumanMouseRatPrimateRabbitSheep |
| Concentration | 1mg/ml (Please refer to the vial label for the specific concentration) |
| Purification | Affinity purified |
| Gene Synonyms | Hsp90aa1 86kDa 89kDa AL024080 AL024147 Hsp86-1 Hsp89 Hsp90 Hspca hsp4 heat shock protein 90 alpha (cytosolic) class A member 1 |
|---|---|
| Target | Heat Shock Protein 86 |
Application
Western blot of mouse HSP 84using
Application
Immunofluorescent analysis of Heat Shock Protein 86 (HSP86, green) in HeLa cells and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a HSP86 polyclonal antibody , at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.
Application
Immunohistochemistry was performed on differentiated colon adenocarcinoma from paraffin-embedded human colon tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA in PBST for 30 minutes at room temperature and then probed with a Heat Shock Protein 86 (Hsp86) polyclonal antibody at a dilution of 1:100 for 1 hour in a humidified chamber (right panel). As a negative control, the primary antibody was eliminated from the staining procedure (left panel). Tissues were washed extensively with PBS/0.025% Tween-20 and endogenous peroxidase activity quenched with Peroxidase Suppressor for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:250 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit. Tissues were counterstained with hematoxylin and prepped for mounting.
Application
Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody at a dilution of 1:1000 overnight at 4ºC on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using ECL Substrate.
Application
Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Application
Western blot analysis of Heat Shock Protein 86 (HSP86) was performed by loading 50ug of the indicated whole cell lysates and 15ul of PageRuler Prestained Protein Ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSP86 polyclonal antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG HRP secondary antibody at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed
Application
Immunohistochemistry was performed on normal deparaffinized Human kidney tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Application
Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing Heat Shock Protein 90 (86) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Application
Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an AP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using Lumi-Phos Substrate.
Application
Western blot analysis of epidermoid carcinoma (A431) cells using Heat Shock Protein 86 Polyclonal Antibody . Decreasing amounts of A431 whole cell lysates were probed with PA3-013 at a dilution of 1:2000 followed by an anti-rabbit IgG secondary antibody and SuperSignal West Pico Chemiluminescent Substrate. Lanes 1 and 2 are control samples of purified, recombinant Hsp86 (rHsp86) and Hsp84 (rHsp84), respectively. Data courtesy of the Innovators Program.| Application Notes | IHC-P: 5 µg/ml: |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution. Aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 1 mg/ml BSA and 0.05% sodium azide. |
