Recombinant Anti-Hsp90 alpha antibody [68/Hsp90] (ab282108)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [68/Hsp90] to Hsp90 alpha
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-Hsp90 alpha antibody [68/Hsp90]
See all Hsp90 alpha primary antibodies -
Description
Mouse monoclonal [68/Hsp90] to Hsp90 alpha -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Wild-type HEK-293T,Jurkat and HeLa, K-562 lysates. ICC: 293T, HeLa cells.
-
General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
68/Hsp90 -
Isotype
IgG1 -
Research areas
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab282108 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB |
1/1000.
|
|
ICC/IF |
1/50.
|
Notes |
---|
WB
1/1000. |
ICC/IF
1/50. |
Target
-
Function
Molecular chaperone. Has ATPase activity. -
Sequence similarities
Belongs to the heat shock protein 90 family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
-
Database links
- Entrez Gene: 3320 Human
- Omim: 140571 Human
- SwissProt: P07900 Human
- Unigene: 525600 Human
- Unigene: 700831 Human
-
Alternative names
- EL52 antibody
- epididymis luminal secretory protein 52 antibody
- Heat shock 86 kDa antibody
see all
Images
-
All lanes : Anti-Hsp90 alpha antibody [68/Hsp90] (ab282108) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : HSP90AA1 KO HEK293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution
Observed band size: 83 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Lanes 1-3: Merged signal (red and green). Green - ab282108 observed at 83KDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.
Lanes 1-2: ab282108 Anti-Hsp90 alpha antibody was shown to react with Hsp90 alpha in HEK293T cells in Western blot. Loss of signal was observed when HSP90AA1 knockout sample was used. Wild-type and
HSP90AA1 knockout samples were subjected to SDS-PAGE.
ab282108 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging. -
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Hsp90 alpha with ab282108 at 1/50 (19.98 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in 293T cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
All lanes : Anti-Hsp90 alpha antibody [68/Hsp90] (ab282108) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
Lane 2 : K-562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Observed band size: 83 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 7 seconds
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Hsp90 alpha with ab282108 at 1/50 (19.98 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in HeLa cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (1)
ab282108 has been referenced in 1 publication.
- Wei J et al. Cucurbitacin B-induced G2/M cell cycle arrest of conjunctival melanoma cells mediated by GRP78-FOXM1-KIF20A pathway. Acta Pharm Sin B 12:3861-3876 (2022). PubMed: 36213538