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Products:Cell Biology >> Proteolysis / Ubiquitin >> Proteasome / Ubiquitin >> Ubiquitin E2s and E3s >> Ubiquitin E2 Enzymes
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If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »Publishing research using ab36980? Please let us know so that we can cite the reference in this datasheet
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - UBCH6/UBE2E1 antibody (ab36980)
Anti-UBCH6/UBE2E1 antibody (ab36980) at 1 µg/ml + HUVEC whole cell lysate at 20 µg
Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 21 kDa
Observed band size : 24 kDa (why is the actual band size different from the predicted?)
Immunoprecipitation - Anti-UBCH6/UBE2E1 antibody (ab36980)
UBCH6/UBE2E1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to UBCH6/UBE2E1 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, Hela 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 ab36980.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 25kDa: hnRNP A1; non specific - 18kDa: We are unsure as to the identity of this extra band.
Immunocytochemistry/ Immunofluorescence - UBCH6/UBE2E1 antibody (ab36980)
ICC/IF image of ab36980 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab36980, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
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