MA3-010
antibody from Invitrogen Antibodies
Targeting: HSP90AA1
FLJ31884, Hsp89, Hsp90, HSP90N, HSPC1, HSPCA
Antibody data
- Antibody Data
- Antigen structure
- References [14]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [5]
- Immunohistochemistry [4]
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Validation data
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- Product number
- MA3-010 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HSP90 alpha Monoclonal Antibody (3B6)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA3-010 detects heat shock protein 90 kDa (HSP90) from human, mouse, rat, and bovine tissues. This antibody does not detect chicken HSP90.
- Antibody clone number
- 3B6
- Concentration
- Conc. Not Determined
Submitted references Heat Shock Protein 90 Involvement in the Development of Idiopathic Epiretinal Membranes.
The distinct agonistic properties of the phenylpyrazolosteroid cortivazol reveal interdomain communication within the glucocorticoid receptor.
Oolemmal proteomics--identification of highly abundant heat shock proteins and molecular chaperones in the mature mouse egg and their localization on the plasma membrane.
Oolemmal proteomics--identification of highly abundant heat shock proteins and molecular chaperones in the mature mouse egg and their localization on the plasma membrane.
Distinct interaction of cortivazol with the ligand binding domain confers glucocorticoid receptor specificity: cortivazol is a specific ligand for the glucocorticoid receptor.
Functional modulation of the glucocorticoid receptor and suppression of NF-kappaB-dependent transcription by ursodeoxycholic acid.
Effect of stages of lactation on the concentration of a 90-kilodalton heat shock protein in bovine mammary tissue.
Mutational analysis of Hsp90 alpha dimerization and subcellular localization: dimer disruption does not impede "in vivo' interaction with estrogen receptor.
Agonist-free transformation of the glucocorticoid receptor in human B-lymphoma cells.
Agonist-free transformation of the glucocorticoid receptor in human B-lymphoma cells.
High levels of non-activated receptors in glucocorticoid-sensitive S49wt mouse lymphoma cells incubated with dexamethasone.
High levels of non-activated receptors in glucocorticoid-sensitive S49wt mouse lymphoma cells incubated with dexamethasone.
ATP-dependent protein refolding activity in reticulocyte lysate. Evidence for the participation of different chaperone components.
ATP-dependent protein refolding activity in reticulocyte lysate. Evidence for the participation of different chaperone components.
Tosi GM, Regoli M, Altera A, Galvagni F, Arcuri C, Bacci T, Elia I, Realini G, Orlandini M, Bertelli E
Investigative ophthalmology & visual science 2020 Jul 1;61(8):34
Investigative ophthalmology & visual science 2020 Jul 1;61(8):34
The distinct agonistic properties of the phenylpyrazolosteroid cortivazol reveal interdomain communication within the glucocorticoid receptor.
Yoshikawa N, Yamamoto K, Shimizu N, Yamada S, Morimoto C, Tanaka H
Molecular endocrinology (Baltimore, Md.) 2005 May;19(5):1110-24
Molecular endocrinology (Baltimore, Md.) 2005 May;19(5):1110-24
Oolemmal proteomics--identification of highly abundant heat shock proteins and molecular chaperones in the mature mouse egg and their localization on the plasma membrane.
Calvert ME, Digilio LC, Herr JC, Coonrod SA
Reproductive biology and endocrinology : RB&E 2003 Feb 14;1:27
Reproductive biology and endocrinology : RB&E 2003 Feb 14;1:27
Oolemmal proteomics--identification of highly abundant heat shock proteins and molecular chaperones in the mature mouse egg and their localization on the plasma membrane.
Calvert ME, Digilio LC, Herr JC, Coonrod SA
Reproductive biology and endocrinology : RB&E 2003 Feb 14;1:27
Reproductive biology and endocrinology : RB&E 2003 Feb 14;1:27
Distinct interaction of cortivazol with the ligand binding domain confers glucocorticoid receptor specificity: cortivazol is a specific ligand for the glucocorticoid receptor.
Yoshikawa N, Makino Y, Okamoto K, Morimoto C, Makino I, Tanaka H
The Journal of biological chemistry 2002 Feb 15;277(7):5529-40
The Journal of biological chemistry 2002 Feb 15;277(7):5529-40
Functional modulation of the glucocorticoid receptor and suppression of NF-kappaB-dependent transcription by ursodeoxycholic acid.
Miura T, Ouchida R, Yoshikawa N, Okamoto K, Makino Y, Nakamura T, Morimoto C, Makino I, Tanaka H
The Journal of biological chemistry 2001 Dec 14;276(50):47371-8
The Journal of biological chemistry 2001 Dec 14;276(50):47371-8
Effect of stages of lactation on the concentration of a 90-kilodalton heat shock protein in bovine mammary tissue.
Watanabe A, Miyamoto T, Katoh N, Takahashi Y
Journal of dairy science 1997 Oct;80(10):2372-9
Journal of dairy science 1997 Oct;80(10):2372-9
Mutational analysis of Hsp90 alpha dimerization and subcellular localization: dimer disruption does not impede "in vivo' interaction with estrogen receptor.
Meng X, Devin J, Sullivan WP, Toft D, Baulieu EE, Catelli MG
Journal of cell science 1996 Jul;109 ( Pt 7):1677-87
Journal of cell science 1996 Jul;109 ( Pt 7):1677-87
Agonist-free transformation of the glucocorticoid receptor in human B-lymphoma cells.
van den Berg JD, Smets LA, van Rooij H
The Journal of steroid biochemistry and molecular biology 1996 Feb;57(3-4):239-49
The Journal of steroid biochemistry and molecular biology 1996 Feb;57(3-4):239-49
Agonist-free transformation of the glucocorticoid receptor in human B-lymphoma cells.
van den Berg JD, Smets LA, van Rooij H
The Journal of steroid biochemistry and molecular biology 1996 Feb;57(3-4):239-49
The Journal of steroid biochemistry and molecular biology 1996 Feb;57(3-4):239-49
High levels of non-activated receptors in glucocorticoid-sensitive S49wt mouse lymphoma cells incubated with dexamethasone.
van den Berg JD, Smets LA, Hutchison KA, van Rooij H, van den Elshout MM
The Journal of steroid biochemistry and molecular biology 1994 Oct;51(1-2):33-40
The Journal of steroid biochemistry and molecular biology 1994 Oct;51(1-2):33-40
High levels of non-activated receptors in glucocorticoid-sensitive S49wt mouse lymphoma cells incubated with dexamethasone.
van den Berg JD, Smets LA, Hutchison KA, van Rooij H, van den Elshout MM
The Journal of steroid biochemistry and molecular biology 1994 Oct;51(1-2):33-40
The Journal of steroid biochemistry and molecular biology 1994 Oct;51(1-2):33-40
ATP-dependent protein refolding activity in reticulocyte lysate. Evidence for the participation of different chaperone components.
Nimmesgern E, Hartl FU
FEBS letters 1993 Sep 27;331(1-2):25-30
FEBS letters 1993 Sep 27;331(1-2):25-30
ATP-dependent protein refolding activity in reticulocyte lysate. Evidence for the participation of different chaperone components.
Nimmesgern E, Hartl FU
FEBS letters 1993 Sep 27;331(1-2):25-30
FEBS letters 1993 Sep 27;331(1-2):25-30
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Heat Shock Protein 90 (HSP90) was performed by loading 50 µg of various whole cell lysates and 15 µL of PageRuler Prestained Protein Ladder (Product # 26616) 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 an HSP90 monoclonal antibody (Product # MA3-010) at a dilution of 1:500 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1), U-87 MG (Lane 2), HEK 293T (Lane 3), HeLa (Lane 4), K-562 (Lane 5) and NIH/3T3 (Lane 6). The blot was probed with Anti-HSP90 alpha Monoclonal Antibody (3B6) (Product # MA3-010, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 80 kDa band corresponding to HSP90 alpha was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing ofHSP90 alpha (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR610567_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of HSP90 alpha was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Cas9 cells transduced with HSP90 alpha Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-HSP90 alpha Monoclonal Antibody (3B6) (Product # MA3-010) using 1:1,000 dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:8,000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toHSP90 alpha (Fig (b)).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HSP90 alpha was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HSP90 alpha Monoclonal Antibody(3B6) (Product # MA3-010) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3B6) (Product # MA3-010) shows staining in NIH-3T3 cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-010) at a dilution of 1:100-1:200 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3B6) (Product # MA3-010) shows staining in HeLa cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-010) at a dilution of 1:100-1:200 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3B6) (Product # MA3-010) shows staining in MCF-7 cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-010) at a dilution of 1:100-1:200 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Heat Shock Protein 90 (HSP90, green) in HeLa 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 (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) an HSP90 monoclonal antibody (Product # MA3-010) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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 1:20 with a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-010) 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on normal 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 1:20 with a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-010) 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on normal 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 1:20 with a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-010) 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on paraffin-embedded human colon cancer 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 (Product # 37525) in PBST for 30 minutes at room temperature and then probed with a Heat Shock Protein 90 (Hsp90) monoclonal antibody (Product # MA3-010) 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 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-mouse IgG-HRP secondary antibody (Product # 31431) at a dilution of 1:250 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Tissues were counterstained with hematoxylin and prepped for mouting. Images were taken on a Zeiss Axiovision microscope at 40X magnification (x1.6 Optovar).