Anti-TPR antibody

• WB

Unknown

Western blot - Anti-TPR antibody (AB70610)

All lanes:

Western blot - Anti-TPR antibody (ab70610) at 0.04 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Lane 4:

293T whole cell lysate at 50 µg

Predicted band size: 267 kDa

Observed band size: 235 kDa,300 kDa

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Exposure time: 30s

• IP

Unknown

Immunoprecipitation - Anti-TPR antibody (AB70610)

Detection of Human TPR by Western Blot of Immunprecipitate. ab70610 at 1µg/ml staining TPR in HeLa whole cell lysates immunoprecipitated using ab70610 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane). Detection : Chemiluminescence with exposure time of 1 second.

All lanes:

Immunoprecipitation - Anti-TPR antibody (ab70610)

Predicted band size: 267 kDa

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• WB

CiteAb

Western blot - Anti-TPR antibody (AB70610)

TPR western blot using anti-TPR antibody ab70610. Publication image and figure legend from Myers, K. N., Barone, G., et al., 2016, Sci Rep, PubMed 27739501.

ab70610 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab70610 please see the product overview.

EBLN1 does not interact with the Cyclin B1-CDK1 complex, but does interact with TPR.(A) Immunoprecipitation of endogenous EBLN1 from HeLa cells probed with either EBLN1 or Cyclin B1 antibodies. Agarose beads incubated with cell extracts and IgG was used as a negative control for non-specific protein binding. Black arrow indicates FLAG-tagged EBLN1 band. (B) Immunoprecipitation (IP) of endogenous Cyclin B1 from asynchronous (upper panel) or mitotic (lower panel) FLAG-EBLN1 expressing HeLa cells probed with the indicated antibodies. The minus tetracycline (−Tet) samples (uninduced expression of FLAG-EBLN1) serve as negative controls for non-specific protein binding. (C) Immunoprecipitation of FLAG-EBLN1 from tetracycline-inducible HeLa cells probed with the indicated antibodies. Minus tetracycline samples serve as negative controls for non-specific protein binding. (D) Upper panel shows EBLN1 and FLAG western blots of eluates from FLAG-EBLN1 expressing tet-inducible HeLa cells. Lower panel shows a SYPRO Ruby stained polyacrylamide gel of FLAG eluates shown in the upper panel. Black arrow indicates FLAG-tagged EBLN1 band. (E) Table showing some of the most prevalent proteins co-immunoprecipitating with FLAG-EBLN1 as determined by proteomic analyses of the eluates shown in (E). The number of unique peptides for each protein is shown for both uninduced and induced (−Tet and + Tet) samples to highlight enrichment in FLAG-EBLN1 eluates (+Tet samples), along with the respective peptide coverage for each protein identified. (F) Indicated western blots on inputs (left panel), GFP immunoprecipitations and TPR immunoprecipitations (right panels) in stable tetracycline-inducible GFP-EBLN1 expressing HeLa cell lines. Arrows highlight GFP-EBLN1 band in each IP. Note that a longer exposure is shown for the TPR IPs compared with the GFP IPs. (G) TPR western blots of immunoprecipitated endogenous TPR from HeLa cells transfected with the indicated siRNA. Arrows highlight TPR isoforms (upper panel) and TPR-specific band in EBLN1 IPs (lower panel), which are reduced in cells transfected with TPR siRNA. Note that the TPR antibody is not capable of recognising endogenous TPR in the input lanes, only purified TPR in the IP lanes.

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