Cells were rinsed in ice-cold phosphate-buffered saline (PBS) and extracts prepared in lysis buffer (50 mM Tris HCl pH 7.5, 1 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 10 mM Na β-glycerophosphate, 50 mM NaF, 5 mM Na Pyrophosphate, 270 mM sucrose and 1% Triton X-100) supplemented with protease inhibitor (Roche) and 1 mM phenylmethylsulfonyl fluoride. Lysates were subjected to SDS-PAGE and transferred to a PVDF membrane (ImmunolonP, Thermo). The following antibodies were used for immunoblotting: anti-rabbit and anti-mouse horseradish peroxidase-conjugated secondary antibodies (1:5000; Pierce); anti-human Hsp90 (1:1000; 610418, BD Biosciences), anti-HIV-Gag (ab9071) and anti-GAPDH (ab9485) 1:2500 from abcam and anti-ADAR1 (14175), anti-MDA5 (5321), anti-phosphoSTAT1 (9167), anti-RIG-I (3743 l), anti-IRF3 (11904), anti-IRF7 (4920) and anti-PKR (12297) all 1:1000 from Cell Signaling. Western blot bands were quantified using Image Studio Software (LI-Cor biosciences).
Anti human hsp90
Manufactured by BD
Sourced in United States
Anti-human Hsp90 is a laboratory reagent used in research applications. It is designed to detect and measure the presence of the heat shock protein 90 (Hsp90) in human samples. Hsp90 is a molecular chaperone involved in the folding and stability of various client proteins. The anti-human Hsp90 reagent can be used in techniques such as Western blotting, immunoprecipitation, and enzyme-linked immunosorbent assay (ELISA) to study the expression and function of Hsp90 in different biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Immunolonp, by Thermo Fisher Scientific (2 mentions)
Ab67820, by Abcam (2 mentions)
Image studio software, by LI COR (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Anti mda5, by Cell Signaling Technology (1 mentions)
Anti rig 1, by Cell Signaling Technology (1 mentions)
Anti hpv16 e7, by Abcam (1 mentions)
Anti phosphorylated irf 7, by Cell Signaling Technology (1 mentions)
Anti adar1, by Cell Signaling Technology (1 mentions)
Anti phospho stat1, by Cell Signaling Technology (1 mentions)
Ab9485, by Abcam (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
4 protocols using anti human hsp90
1
Immunoblotting Assay for Viral and Cellular Proteins
2
Western Blot Analysis of Cell Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell extracts were prepared for Western blot protein analysis as described before [22 (link)]. Briefly, cells were rinsed in ice-cold phosphate-buffered saline (PBS) and extracts prepared in lysis buffer (50 mM Tris HCl pH 7.5, 1 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 10 mM Na β-glycerophosphate, 50 mM NaF, 5 mM Na Pyrophosphate, 270 mM sucrose and 1% Triton X-100) supplemented with protease inhibitor (Roche, Basel, Switzerland) and 1 mM phenylmethylsulfonyl fluoride. Lysates were subjected to SDS-PAGE and transferred to a PVDF membrane (ImmunolonP, Thermo, Waltham, MA, USA). The following antibodies were used for immunoblotting: anti-rabbit and anti-mouse horseradish peroxidase-conjugated secondary antibodies (1:5000; Pierce, Waltham, MA, USA; anti-human Hsp90 (BD Biosciences, Franklin Lakes, NJ, USA; 610418), anti-SAMHD1 (1:2500; ab67820, Abcam, Cambridge, UK) and anti-RRM2 (1:1000; ab115701, Abcam) and anti-phospho-CDK2 (Thr160; 2561), anti-CDK2 (2546), anti-CDK1 (77055) anti-Rb (9309) anti-phospho-Rb (Ser807/811, 9308) anti-E2F1 (3742), anti-phospho-SAMHD1 Thr592 (15038) (all 1:1000; Cell Signaling Technologies, Danvers, MA, USA).
3
Western Blot Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein expression was evaluated by Western blot as described before37 (link). The following antibodies were used for immunoblotting: anti-rabbit and anti-mouse horseradish peroxidase-conjugated secondary antibodies (1:5000; Pierce); anti-human Hsp90 (1:1000; 610418, BD Biosciences), anti-ADAR1 (1:1000; 14175; Cell Signaling); anti-MDA5 (1:500; 5321; Cell Signaling); anti-RIG-I (1:1000; 3743; Cell Signaling); anti-phosphoSTAT1 (1:1000; 9167; Cell Signaling); anti-phosphorylated IRF7 (1:1000; 12390; Cell Signaling); anti-GAPDH (1:1000; ab9485; abcam); and anti-HPV16 E7 (1:1000; ab82601; Abcam).
4
Immunoblotting of Phosphorylated SAMHD1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Treated cells were rinsed in ice-cold PBS, extracts were prepared in lysis buffer (50 mM Tris HCl pH 7.5, 1 mM EDTA, 1 mM EGTA, 1 mM NaV 3 O 4 , 10 mM sodium b-glycerophosphate, 50 mM NaF, 5 mM sodium pyrophosphate, 270 mM sucrose and 1% Triton X-100) supplemented with protease inhibitor cocktail (Roche) and 1 mM phenylmethylsulfonyl fluoride. Samples were electrophoresed in SDS-polyacrylamide gels containing 6e10% acrylamide, and blotted onto nitrocellulose membranes. Blocked membranes were incubated overnight at 4 C with the following antibodies: anti-rabbit and anti-mouse horseradish peroxidaseconjugated secondary antibodies (1:5000; Pierce); anti-human Hsp90 (1:1000; 610418, BD Biosciences); anti-SAMHD1 (1:2500; ab67820, Abcam); anti-phospho(Thr)-Pro (ref. 23,21) Actin (1:5000; ref. A5316, . Anti-phospho-SAMHD1 Thr592 (pSAMHD1 T592) was obtained by immunization of rabbit using a phosphorylated peptide as described before (White et al., 2013b) . After washing, the membranes were incubated with a secondary conjugated horseradish peroxidase antibody for 1 h at room temperature and then revealed with SuperSignal West Pico Chemiluminescent Substrate (Pierce Chemical, Rockford, IL, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!