Anti-Lin28A antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Lin28A antibody (AB63740)

ab63740 staining Lin28A in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab63740 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IP

Unknown

Immunoprecipitation - Anti-Lin28A antibody (AB63740)

Lin28A was immunoprecipitated using 0.5mg E14Tg2a whole cell extract, 5μg of Rabbit polyclonal to Lin28A 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 : Lin28A.

All lanes:

Immunoprecipitation - Anti-Lin28A antibody (ab63740)

Predicted band size: 23 kDa

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• WB

Project5080****

Western blot - Anti-Lin28A antibody (AB63740)

The 30 kDa band observed is comparable to molecular weights seen with other commercially available antibodies to Lin28A.

All lanes:

Western blot - Anti-Lin28A antibody (ab63740) at 1 µg/mL

Lane 1:

MEL-1 (Human embryonic stem cell, male cell line) Whole Cell Lysate (<a href='/en-us/products/unavailable/mel-1-human-embryonic-stem-cell-male-cell-line-whole-cell-lysate-ab27198'>ab27198</a>) at 10 µg

Lane 2:

F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate (<a href='/en-us/products/unavailable/f9-mouse-embryonic-carcinoma-cell-line-whole-cell-lysate-ab27193'>ab27193</a>) at 10 µg

Lane 3:

E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 µg

Lane 4:

Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 23 kDa

Observed band size: 30 kDa

false

• WB

Ap16151****

Western blot - Anti-Lin28A antibody (AB63740)

This 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.

All lanes:

Western blot - Anti-Lin28A antibody (ab63740) at 1 µg/mL

Lane 1:

HUES7 (Human embryonic stem cell line) Whole Cell Lysate at 10 µg

Lane 2:

F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 23 kDa

Observed band size: 30 kDa

true

Exposure time: 90s

• sELISA

Unknown

Sandwich ELISA - Anti-Lin28A antibody (AB63740)

Standard Curve for Lin28A (Analyte : (ab89225) Lin28 protein (His tag) (ab89225)); dilution range 1 pg/ml to 1 ug/ml using Capture Antibody (ab76369) Mouse monoclonal [6D1F9] to Lin28A (ab76369) at 0.2 ug/ml and Detector Antibody (ab63740) Rabbit polyclonal to Lin28A (ab63740) at 0.1 ug/ml.