PA1-10023

antibody from Invitrogen Antibodies

Targeting: UBC

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-10023

Provider

Invitrogen Antibodies

Product name

Ubiquitin Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Purifed from natural sources

Description

PA1-10023 was made against purified bovine blood derived ubiquitin coupled to keyhole limpet hemocyanin with glutaraldehyde.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ubiquitin in HEK293 cell lysates. The lysate was subjected to electrophoresis on a 4-20% SDS-PAGE gel, then electrophoretically transferred to PVDF membranes. Ubiquitin on the membrane was detected using a polyclonal antibody (Product # PA1-10023) at a dilution of 1:5,000 as seen in green, and a beta-Tubulin monoclonal antibody at a dilution of 1:10,000 used as a loading control as seen in red. 1) protein standard (red), 2) cells maintained in normal medium, 3) cells treated with proteasome inhibitor lactacystin (Lc) at 10µM for 16 hours. The smear detected above the 200 kDa standard represents accumulations of ubiquitinated proteins in the Lc treated cells. The prominent band at ~8 kDa corresponds to monoubiquitin.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Ubiquitin in HeLa cells. Cells were stained with a ubiquitin polyclonal antibody (Product # PA1-10023) at a dilution of 1:1,000 as seen in red, stained with a Vimentin polyclonal antibody (Product # PA1-10003) at a dilution of 1:10,000 as seen in green, and with DAPI staining the nuclear DNA as seen in blue. A) Control HeLa cells maintained in normal medium, B) HeLa cells treated with 10µM of the proteasome inhibitor lactacystin (Lc) for 24 hours. Proteasomal inhibition leads to formation of strongly ubiquitin positive cytoplasmic inclusions. Note the diffuse cytoplasmic ubiquitin staining in control cells and the well-defined ubiquitin positive inclusions in the Lc treated cells.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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