Anti-Hsp90 alpha抗体
参阅全部 Hsp90 alpha 一抗
兔多克隆抗体to Hsp90 alpha
Rabbit
Detects Heat Shock Protein 86 (HSP 86). This antibody does not detect HSP 84.
适用于: ICC/IF, WB, IP, IHC-Pmore details
与反应: Mouse, Rat, Human, African green monkey
Synthetic peptide corresponding to Mouse Hsp90 alpha aa 2-12 (N terminal).
Sequence:
PEETQTQDQPM
WB: HEK-293T, HeLa, K562, A431, HepG2, COS-7, NIH/3T3, NRK whole cell lysate. ICC/IF: HeLa, NIH/3T3, whole cell. IHC-P: Human colon adenocarcinoma, breast carcinoma, tonsil, kidney tissue. IP: HeLa whole cell lyaste.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
Heat shock proteins (HSP) are expressed in response to various biological stresses, including heat. HSP 90 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. HSP 90 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 HSP 90 exists as two forms, HSP 84 and HSP 86, coded by related but separate genes, with 86% homologous amino acid sequences. These forms are analogous to the two forms of human HSP 90, HSP 89 beta and HSP 89 alpha. In an unstressed mouse fibroblast, the basal level of HSP 84 is found to be double that of HSP 86. However, after heat shock, HSP 86 shows a greater increase. Studies also suggest that upon cellular differentiation, the level of HSP 86, but not HSP 84, decreases. HSP 84 and HSP 86, 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.
多克隆
IgG
Abpromise™承诺保证使用ab2928于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | 1/50 - 1/200. | |
| WB | 1/500 - 1/2000. Detects a band of approximately 86 kDa. | |
| IP | Use at an assay dependent concentration. Use at 2 μg. Immunoprecipitation experiments with this antibody suggest that HSP90 alpha exists primarily as homodimers in HeLa cells. This antibody is capable of precipitating HSP90 alpha that is complexed with other proteins such as the aryl hydrocarbon (Ah) receptor. Read More | |
| IHC-P | Use a concentration of 5 µg/ml. |
Entrez Gene: 3320 Human
Entrez Gene: 15519 Mouse
Omim: 140571 Human
SwissProt: P07900 Human
SwissProt: P07901 Mouse
Unigene: 525600 Human
Unigene: 700831 Human
Unigene: 1843 Mouse
Unigene: 341186 Mouse
Unigene: 119867 Rat
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Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody (ab2928)
Immunocytochemistry analysis of HeLa cells labeling HSP90 alpha with ab2928 at 5ug/mL in 0.1% BSA, incubated at 4ºC overnight. Cells were 70% confluent log phase. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. Cells were then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 at 1/2000, for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI. F-actin (Panel c: Red) was stained with Rhodamine Phalloidin 1/300). Panel d represents the merged image showing cytoplasm and weak Nucleus localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Western blot - Anti-Hsp90 alpha antibody (ab2928)
All lanes : Anti-Hsp90 alpha antibody (ab2928) at 1/1000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 6 : COS-7 (African green monkey kidney fibroblast-like cell line) whole cell lysate
Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 8 : NRK (Rat kidney normal tissue) whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : Goat anti-rabbit IgG HRP secondary antibody at 1/20000 dilution
Western blot analysis of HSP90 alpha was performed by loading samples 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 ab2928 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a secondary antibody for at least 1 hour. Chemiluminescent detection was performed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
ab2928 labelling Hsp90 alpha in Human colon adenocarcinoma tissue sections by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Tissues were blocked in 3% BSA in PBST for 30 minutes at room temperature. Tissue sections were incubated with the primary antibody (1:100) for 1 hour. A HRP-conjugated goat anti-rabbit IgG (1:250) was used as the secondary antibody, followed by colorimetric detection using Metal Enhanced DAB Substrate Kit. Tissues were counterstained with hematoxylin and prepped for mouting. Images were taken at 40X magnification.
Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody (ab2928)
Immunocytochemistry/Immunofluorescence analysis of HSP90 alpha (green) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and NIH/3T3 (Mouse embryo fibroblast cell line) cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were incubated with ab2928 at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat-anti-rabbit IgG secondary antibody (1:400) for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.
Western blot - Anti-Hsp90 alpha antibody (ab2928)
All lanes : Anti-Hsp90 alpha antibody (ab2928) at 1/2000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A549 cell lysate
Lane 3 : LNCaP cell lysate
Lane 4 : NIH/3T3 cell lysate
Lane 5 : Mouse testis tissue lysate
Lane 6 : Rat testis tissue lysate
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant, HRP at 1/4000 dilution
Observed band size: 80 kDawhy is the actual band size different from the predicted?
Detected by chemiluminescence
Immunoprecipitation - Anti-Hsp90 alpha antibody (ab2928)
Immunoprecipitation of HSP90 alpha was performed on HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. Antigen-antibody complexes formed by incubating 500ug whole cell lysate with 2ug of ab2928 overnight on a rocking platform at 4°C. The immune complexes were captured on 50ul Protein A/G Plus Agarose, washed extensively, and eluted with buffer. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab2928 at a dilution of 1:1000 overnight rotating at 4°C. The membrane was washed in TBST, and probed with detection reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
Immunohistochemistry was performed on both normal and 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) ab2928 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue. 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) ab2928 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 alpha antibody (ab2928)
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human kidney tissue. 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) ab2928 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.
