Western blot analysis of Heat Shock Protein 86 (Hsp86) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Hsp86 polyclonal antibody (Product # PA3-013) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31466) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using ECL Substrate (Product # 32106).
|Tested species reactivity||Rabbit|
|Published species reactivity||Not Applicable|
|Host / Isotype||Goat / IgG|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.6, with 15mg/ml BSA, 50mM sucrose, proprietary stabilizer|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||1:5,000 - 1:100,000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:5,000-1:100,000|
|Immunoprecipitation (IP)||1:500 - 1:5,000|
|Western Blot (WB)||1:5,000 - 1:200,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Product # 31466 has been successfully used in Western blot, IHC and IP applications.
Product # 31466 reacts with the heavy chains of rabbit IgG and with the light chains common to most rabbit immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species and with SuperBlock® Blocking Buffers.
Store product at 4°C until opened. To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C.
Reconstitute with 0.2 ml of distilled water (1 mg/ml after restoration).
Thermo Scientific Anti-Rabbit secondary antibodies are affinity-purified antibodies with well-characterized specificity for rabbit immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Sirtuin1 promotes osteogenic differentiation through downregulation of peroxisome proliferator-activated receptor ¿ in MC3T3-E1 cells.
31466 was used in western blot to investigate the molecular mechanism of Sirt1 in osteogenic differentiation
|Qu B,Ma Y,Yan M,Gong K,Liang F,Deng S,Jiang K,Ma Z,Pan X||Biochemical and biophysical research communications (478:439)||2016|
Inhibition of autophagy using 3-methyladenine increases cisplatin-induced apoptosis by increasing endoplasmic reticulum stress in U251 human glioma cells.
31466 was used in western blot to ascertain the role of autophagy in cisplatin chemotherapy
|Zhang R,Wang R,Chen Q,Chang H||Molecular medicine reports (12:1727)||2015|
|Not Applicable||Not Cited||
Use of the disulfiram/copper complex for breast cancer chemoprevention in MMTV-erbB2 transgenic mice.
31466 was used in western blot to test the effect of disulfiram/copper complex on breast cancer cells
|Yang Y,Deng Q,Feng X,Sun J||Molecular medicine reports (12:746)||2015|
Distribution of vitellogenin in zebrafish (Danio rerio) tissues for biomarker analysis.
31466 was used in western blot to assess the expression of vitellogenin and its receptor in zebrafish
|Zhong L,Yuan L,Rao Y,Li Z,Zhang X,Liao T,Xu Y,Dai H||Aquatic toxicology (Amsterdam, Netherlands) (149:1)||2014|
Negative pressure wound therapy promotes vessel destabilization and maturation at various stages of wound healing and thus influences wound prognosis.
31466 was used in immunohistochemistry - paraffin section to investigate the effect of negative pressure wound therapy on angiogenesis and vessel maturation
|Ma Z,Shou K,Li Z,Jian C,Qi B,Yu A||Experimental and therapeutic medicine (11:1307)||2016|