Anti-Lin28 antibody (ab63740)
Key features and details
- Rabbit polyclonal to Lin28A
- Suitable for: IP, Sandwich ELISA, ICC/IF, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Lin28A antibody
See all Lin28A primary antibodies -
Description
Rabbit polyclonal to Lin28A -
Host species
Rabbit -
Tested applications
Suitable for: IP, Sandwich ELISA, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Horse, Pig, Chimpanzee
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- This antibody gave a positive signal in the following Whole Cell Lysates: HUES-7 MEL-1 F9 ES-E14TG2a Mouse EG
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Images
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Western blot - Anti-Lin28 antibody (ab63740)All lanes : Anti-Lin28 antibody (ab63740) at 1 µg/ml
Lane 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 Lysate
Lane 4 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
The 30 kDa band observed is comparable to molecular weights seen with other commercially available antibodies to Lin28. -
Immunocytochemistry/ Immunofluorescence - Anti-Lin28 antibody (ab63740)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). -
Western blot - Anti-Lin28 antibody (ab63740)All lanes : Anti-Lin28 antibody (ab63740) at 1 µg/ml
Lane 1 : HUES7 (Human embryonic stem cell line) Whole Cell Lysate
Lane 2 : F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate
Lane 3 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?
Exposure time: 90 secondsThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab63740 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Sandwich ELISA - Anti-Lin28 antibody (ab63740)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. -
Immunoprecipitation - Anti-Lin28 antibody (ab63740)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.
