Anti-Lin28 antibody

• Supplier: Abcam
• Catalog: ab63740
• Prices: $392.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IP ELISA ICC/IF ICC/IF WB
• Species Reactivities: Mouse, Human, Rat, Horse, Pig, Chimpanzee
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Lin28
• Description: Rabbit Polyclonal
• Gene: LIN28A
• Conjugate: Unconjugated

Product images

ICC/IF image of ab63740 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab63740, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).

All lanes : Anti-Lin28 antibody (ab63740) at 1 µg/mlLane 1 : MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (ab27198)Lane 2 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (ab27193)Lane 3 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell LysateLane 4 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionPerformed under reducing conditions.

Standard Curve for Lin28 (Analyte: Lin28 protein (His tag) (ab89225)); dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [6D1F9] to Lin28 (ab76369) at 0.2 ug/ml and Detector Antibody Rabbit polyclonal to Lin28 (ab63740) at 0.1 ug/ml.

Lin28 was immunoprecipitated using 0.5mg E14Tg2a whole cell extract, 5µg of Rabbit polyclonal to Lin28 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).The antibody was incubated under agitation with Protein G beads for 10min, E14Tg2a whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab63740.Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.Band: 30kDa: Lin28.