RPMI1640 (Sigma), Antibiotic antimycotic solution (Sigma),Fetal bovine serum (FBS) Gibco BRL, Pure link RNA extraction kit (Invitrogen Life Technologies), Maxima SYBR Green/ROX qPCR Master Mix(Fermentas), RevertAid Premium first strand cDNA synthesis kit(Fermentas), IPG strips (GE Healthcare), Urea, ThioUrea, CHAPS, EDTA, PMSF,dithiothreitol (DTT), Protease inhibitor cocktail, Iodoacetamide, Bradfoerd, Silver nitrate, Ponceau, BSA, poly-L lysine and paraformaldehyde were purchased from Sigma Aldrich. Passive Lysis Buffer (Promega), PVDF membrane (Millipore), Luminata forte Western HRP substrate (Millipore), anti alpha-tubulin monoclonal antibody (Invitrogen Life Technologies), DAPI (Invitrogen Life Technologies), Primary antibodies- anti-Cyclophilin A antibody (ab58144), anti-eIF5A antibody [EP527Y] (ab32407), anti-ENO1 antibody (ab85086), anti-OTUB1 antibody (ab98280), anti-RhoGDI antibody [1F2] (ab118159), and anti-VDAC1/Porin antibody—N-terminal (ab135585) were purchased from abcam, UK.
Ab85086
Manufactured by Abcam
Sourced in United Kingdom
Ab85086 is a recombinant protein product manufactured by Abcam. It is a tool used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Urea, by GE Healthcare (1 mentions)
Thiourea, by GE Healthcare (1 mentions)
Ab32407, by Abcam (1 mentions)
Ponceau, by Merck Group (1 mentions)
Ipg strip, by GE Healthcare (1 mentions)
Maxima sybr green rox qpcr master mix, by Thermo Fisher Scientific (1 mentions)
Purelink rna extraction kit, by Thermo Fisher Scientific (1 mentions)
Revertaid premium first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Chaps, by GE Healthcare (1 mentions)
Luminata forte western hrp substrate, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Iodoacetamide, by Merck Group (1 mentions)
3 protocols using ab85086
1
Proteomic analysis of cell lines
2
Western Blot Analysis of Protein Expression
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Total protein was extracted from indicated cells or tumor tissues using lysis buffer (2% SDS, 100 mM DTT, 10 mM Tris (pH 6.8) and 10% glycerol). Tumor tissues in lysis buffer were sonicated at a frequency of 70 Hz for 180 s. Then they were centrifuged at 12000 rpm for 5 min and boiled at 98°C for 10 min. The protein was separated on a 12% or 15% SDS-PAGE gel and transferred to polyvinylidenefluoride membrane (PVDF; Millipore, U.S.A.). The membranes were blocked with 5% silk milk at room temperature for 1 h and incubated with primary antibodies at 4°C overnight. Antibody against ENO1 (ab85086, 1:1000), cleaved caspase 3 (ab2303) and 9 (ab2324) was from Abcam (U.K.). Antibodies against cyclin D1 (sc-8396, 1:500), E1 (sc-377100, 1:500), p21 (cst2947, 1:1000), p27 (cst3686, 1:1000), caspase 3 (cst9662, 1:1000) and 9 (cst9502, 1:1000) were from Cell Signaling (U.S.A.) and Santa Cruz Biotech (U.S.A.). Antibody against β-catenin (cst8480, 1:1000) was from Cell Signaling (U.S.A.). GAPDH antibody was from ABclonal (AC033, 1:3000) (U.S.A.). All the secondary antibodies (sc-2537 and sc-2005, 1:5000) were from Santa Cruz Biotech (U.S.A.).
3
Immunohistochemical Profiling of Pancreatic Cancer
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The commercial tissue microarrays were constructed by Shanghai Biochip Co. Ltd., as described previously 15 (link). Briefly, the tissue microarrays including 100 pancreatic cancer patients and 80 adjacent normal tissues were prepared from archival formalin-fixed, paraffin embedded tissue blocks. A representative tumor area was carefully selected from a H&E-stain section. For all the specimens, clinicopathological information (age, gender, and pathology, differentiation, and TNM stage) and Follow-up information were available. Standard Avidin-biotin complex peroxidase immunohistochemical staining was performed. Briefly, after deparaffinizationin xylene and graded alcohols, heated antigen retrieval was done in citrate buffer (10mmol/L pH 6.0) by water-bath kettle heating for 30min. Endogenous peroxidase was blocked in 0.3% hydrogen peroxide for 10 min. Nonspecific binding was blocked by incubation in 10% normal animal serum for 10min. Sections were incubated at 4°C for 24 h with primary antibodies including polyclonal antibody against anti-alpha-enolase (ab85086, Abcam), Anti-p53 antibody (ab28, Abcam) and anti-Ki67 (ab833, Abcam). Next, biotinylated secondary antibodies and horseradish peroxidase labeled avidin were incubated with samples. Color was developed using the DAB method.
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