Anti-UBR7 antibody [EPR26170-67]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling UBR7 with ab305239 at 1/500 (0.942 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Nuclear staining in human testis. The section was incubated with ab305239 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling UBR7 with ab305239 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC

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Immunocytochemistry - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling UBR7 with ab305239 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in MCF7 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-UBR7 antibody [EPR26170-67] (AB305239)

UBR7 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate 10 μg with ab305239 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305239 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate 10 μg Lane 2 : ab305239 IP in MCF7 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305239 in MCF7 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds The bands beneath the target band (55 kDa) are likely to be degraded target fragments

All lanes:

Immunoprecipitation - Anti-UBR7 antibody [EPR26170-67] (ab305239) at 1/30 dilution

All lanes:

MCF7 (human breast adenocarcinoma epithelial cell)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 55 kDa

false

Exposure time: 32s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling UBR7 with ab305239 at 1/500 (0.942 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Nuclear staining in mouse colon. The section was incubated with ab305239 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling UBR7 with ab305239 at 1/500 (0.942 µg/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Nuclear staining in rat colon. The section was incubated with ab305239 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling UBR7 with ab305239 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling UBR7 with ab305239 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-UBR7 antibody [EPR26170-67] (AB305239)

UBR7 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 μg with ab305239 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305239 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 μg Lane 2 : ab305239 IP in NIH/3T3 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305239 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 24 seconds The bands beneath the target band (55 kDa) are likely to be degraded target fragments

All lanes:

Immunoprecipitation - Anti-UBR7 antibody [EPR26170-67] (ab305239) at 1/30 dilution

All lanes:

NIH/3T3 (mouse embryonic fibroblast)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 55 kDa

false

Exposure time: 24s

• WB

Supplier Data

Western blot - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The bands beneath the target band (55 kDa) are likely to be degraded target fragments Exposure time : 3 minutes

All lanes:

Western blot - Anti-UBR7 antibody [EPR26170-67] (ab305239) at 1/1000 dilution

Lane 1:

T-47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 3:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 4:

Human testis tissue lysate at 20 µg

Lane 5:

Mouse testis tissue lysate at 20 µg

Lane 6:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 48 seconds

All lanes:

Western blot - Anti-UBR7 antibody [EPR26170-67] (ab305239) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell)transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

MCF7 transfected with siRNA specifically targeti UBR7, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-UBR7 antibody [EPR26170-67] (AB305239)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : 3 minutes

All lanes:

Western blot - Anti-UBR7 antibody [EPR26170-67] (ab305239) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 55 kDa

false

Exposure time: 3min