Anti-Hsp90 alpha 抗体 (ab2928)
Key features and details
- Rabbit polyclonal to Hsp90 alpha
- Suitable for: ICC/IF, WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human, African green monkey
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-Hsp90 alpha antibody
Hsp90 alpha 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Hsp90 alpha -
由来種
Rabbit -
特異性
Detects Heat Shock Protein 86 (HSP 86). This antibody does not detect HSP 84. -
アプリケーション
適用あり: ICC/IF, WB, IP, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human, African green monkey -
免疫原
Synthetic peptide corresponding to Mouse Hsp90 alpha aa 2-12 (N terminal).
Sequence:PEETQTQDQPM
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ポジティブ・コントロール
- 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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特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
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精製度
Immunogen affinity purified -
一次抗体 備考
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. -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab2928の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
1/50 - 1/200.
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WB |
1/500 - 1/2000. Detects a band of approximately 86 kDa.
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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. |
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IHC-P |
Use a concentration of 5 µg/ml.
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特記事項 |
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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. |
IHC-P
Use a concentration of 5 µg/ml. |
ターゲット情報
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機能
Molecular chaperone. Has ATPase activity. -
配列類似性
Belongs to the heat shock protein 90 family. -
翻訳後修飾
ISGylated. -
細胞内局在
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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参照データベース
- Entrez Gene: 3320 Human
- Entrez Gene: 15519 Mouse
- Entrez Gene: 299331 Rat
- Omim: 140571 Human
- SwissProt: P07900 Human
- SwissProt: P07901 Mouse
- SwissProt: P82995 Rat
- Unigene: 525600 Human
see all -
別名
- EL52 antibody
- epididymis luminal secretory protein 52 antibody
- Heat shock 86 kDa antibody
see all
画像
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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.
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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 dilutionWestern 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.
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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.
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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.
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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 kDa why is the actual band size different from the predicted?Detected by chemiluminescence
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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.
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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.
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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.
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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (38)
ab2928 は 38 報の論文で使用されています。
- Sharma S et al. Unraveling the Mechanism of Epichaperome Modulation by Zelavespib: Biochemical Insights on Target Occupancy and Extended Residence Time at the Site of Action. Biomedicines 11:N/A (2023). PubMed: 37892973
- Roychowdhury T et al. Use of Native-PAGE for the Identification of Epichaperomes in Cell Lines. Methods Mol Biol 2693:175-191 (2023). PubMed: 37540435
- Rodina A et al. Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation. Nat Commun 14:3742 (2023). PubMed: 37353488
- Ma B et al. Albumosomes formed by cytoplasmic pre-folding albumin maintain mitochondrial homeostasis and inhibit nonalcoholic fatty liver disease. Signal Transduct Target Ther 8:229 (2023). PubMed: 37321990
- Xinxin P et al. Exploring the mechanism of hirudin in the treatment of diabetic kidney disease using network pharmacology combined with molecular docking verification. J Tradit Chin Med 42:586-594 (2022). PubMed: 35848975