Anti-YB1 antibody (ab12148)
Key features and details
- Rabbit polyclonal to YB1
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-YB1 antibody
See all YB1 primary antibodies -
Description
Rabbit polyclonal to YB1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human YB1 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab12411) -
General notes
YB1 has a predicted band size of 36kDa. According to Evdolimova (1995) YB1 migrates by SDS-PAGE at 50kDa, which may be due to post-translational modification. YB1 is primarily detectable in the cytoplasm without any clear signal in nucleoli.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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-YB1 antibody (ab12148)All lanes : Anti-YB1 antibody (ab12148) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 3 : Jurkat (Human) Whole Cell Lysate
Lane 4 :T47D whole cell lysate (ab14899)
Lane 5 : MDA-MB-231 (Human breast adenocarcinoma 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: 36 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab12148 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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Immunocytochemistry/ Immunofluorescence - Anti-YB1 antibody (ab12148)ICC/IF image of ab12148 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 5 min) and incubated with the antibody (ab12148, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT™ FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Western blot - Anti-YB1 antibody (ab12148)Anti-YB1 antibody (ab12148) at 1 µg/ml + HEK293 Whole Cell Lysate Transiently Overexpressing YB1 at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
YB1 has a predicted band size of 36kDa based on its primary sequence (SwissProt). According to Evdolimova (1995) YB1 migrates by SDS-PAGE at 50kDa, which may be due to post-translational modification -
Immunocytochemistry/ Immunofluorescence - Anti-YB1 antibody (ab12148)ICC/IF image of ab12148 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 5 min) and incubated with the antibody (ab12148, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT™ FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Western blot - Anti-YB1 antibody (ab12148)All lanes : Anti-YB1 antibody (ab12148) at 1.4 µg/ml
Lane 1 : HeLa Nuclear lysate
Lane 2 : HeLa Whole cell lysate
Lane 3 : MCF-7 cell lysate
Lane 4 : Jurkat whole cell lysate
Lane 5 : HEK293 Whole cell lysate
Lane 6 : HeLa Nuclear lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 7 : HeLa Whole cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 8 : MCF-7 cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 9 : Jurkat whole cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 10 : HEK293 whole cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Predicted band size: 36 kDa
Observed band size: 36,50 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/ Immunofluorescence - Anti-YB1 antibody (ab12148)ICC/IF image of ab12148 stained HeLa 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 (ab12148, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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) at a concentration of 1.43µM.
