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Primary Antibodies  >  Cancer Research Antibodies
 

HSP 90 K294 Antibody

Rabbit Polyclonal
Code

600-401-981
Size

100 µg
Price

$309.00
Availability

Ships next business day.
Order Offline:
Phone: 800.656.7625
E-mail: orders@rockland-inc.com
Fax: 610.367.7825
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Rockland's affinity purified anti-Hsp90 acetyl K294 antibody was used at 20 µg/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate nuclear and granular cytoplasmic positive staining in human prostate carcinoma at 40X.  Tissue was formalin-fixed and paraffin embedded.  The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.  Personal Communi-cation, Tina Roush, LifeSpanBiosciences, Seattle, WA.
Western blot using Rockland's Affinity Purified anti-Hsp90 acetyl K294 antibody shows detection of a band at ~90 kDa corresponding to Hsp90 in an SkBr3 cell lysate (arrowhead) after treatment with Trichostatin A (an HDAC inhibitor).  Western blotting results do not definitively demonstrate the acetyl K294 specificity of this reagent because similar staining is seen in the control lane (no treatment with TSA).  Immunoprecipitation with anti-Hsp90 was performed prior to western blotting with anti-Hsp90 acetyl K294.
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Synonyms: Heat shock protein HSP 90-alpha Gene names Name: HSP90AA1 Synonyms: HSP90A, HSPCA

HSP 90 K294 Antibody Properties

Description
Anti-Heat Shock Protein HSP 90-alpha acetyl specific K294 (RABBIT) Antibody - 600-401-981
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human, mouse, rat
Clonality
Polyclonal
Application
ELISA : 1:2,500 - 1:10,000
Western Blot : 1:500 - 1:2,500
Immunochemistry: 5-10 µg/ml
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
HSP90AA1
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Lot#  

HSP 90 K294 Antibody Description

Background
This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. Hsp90 is a member of the heat shock protein 90 family, the members of which are highly conserved between isoforms and species. Hsp90 functions as a molecular chaperone and has ATPase activity.  Hsp90 is a cytoplasmic protein that forms a homodimer in vivo and interacts with TOM34, AHSA1, HDAC6 and SMYD3.  Several signal transduction pathways depend on Hsp90 function, including pathways involving erbB2, hypoxia sensitivity (Hif1 alpha), and steroid hormone receptors (for example, androgen, progesterone, glucocorticoid, and aryl-hydrocarbon).  Recent reports show that Hsp90 from tumor cells has increased sensitivity to small molecule inhibitors (for example, 17AAG).  The mechanism of the differential sensitivity of Hsp90 from normal versus tumor cells is unknown, although mutation has been ruled out.  One possible mechanism may be differences in post-translational modification of tumor Hsp90.  K294 was found to be acetylated in purified Hsp90 from SkBr3 cells, a breast cancer cell line.
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids surrounding K294 of human Hsp90.
Immunogen Type
Peptide
Storage Condition
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and western blot.  This antibody reacts strongly with the acetylated form of the immunizing peptide and shows minimal reactivity with the non-acetylated form when tested by ELISA.  To date, western blotting shows equivalent staining of lysates either treated or untreated with Trichostatin A (an HDAC inhibitor).  Therefore, western blotting results are not definitive for demonstrating the specificity of this reagent.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 90 kDa in size corresponding to Hsp90 protein by western blotting in the appropriate cell lysate or extract.
Purity/Specifity
This affinity purified antibody is directed against human Hsp90 protein acetylated at K294. The product was affinity purified from monospecific antiserum by immunoaffinity chromatography using acetyl-peptide coupled to agarose beads followed by solid phase adsorption against non-acetyl peptide.  While ELISA data show strong reactivity with the acetylated form of the immunizing peptide and minimal reactivity with the non-acetylated form, to date western blotting data are not definitive for acetyl K294 specificity as blots show equivalent staining of lysates from cells either treated or untreated with Trichostatin A (an HDAC inhibitor).  A BLAST analysis was used to suggest cross-reactivity with Hsp90 from human, mouse, rat, monkey, chicken and Drosophila based on 100% homology with the immunizing sequence.  Reactivity of this antibody with Hsp90 from other species is unknown.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-268

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF
Reacts With
human
Catalog Number
600-401-879

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
610-4302

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
611-1302

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry