Anti-UBE2C antibody [EPR23165-31] (ab252940)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23165-31] to UBE2C
- Suitable for: WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-UBE2C antibody [EPR23165-31]
See all UBE2C primary antibodies -
Description
Rabbit monoclonal [EPR23165-31] to UBE2C -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF MouseHumanIHC-P MouseRatHumanIP HumanWB MouseHuman -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: T-47D, SW480, NIH:OVCAR-3, HeLa, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. Mouse and rat spleen tissue lysate. IHC-P: Human gastric cancer tissue and colon tissue. Mouse and rat colon tissue. ICC/IF: SW480 and RAW 264.7 cells. IP: SW480 and NIH:OVCAR-3 whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23165-31 -
Isotype
IgG -
Research areas
Images
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Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling UBE2C with ab252940 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasm and nuclear staining in proliferative cells of human gastric cancer is observed (PMID:15208666, 27349176). The section was incubated with ab252940 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling UBE2C with ab252940 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in partial RAW 264.7 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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: ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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All lanes : Anti-UBE2C antibody [EPR23165-31] (ab252940) at 1/1000 dilution
Lane 1 : Mouse spleen lysate at 20 µg
Lane 2 : Rat spleen lysate at 20 µg
Lane 3 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 10 µg
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg
Lane 5 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 20 kDa
Observed band size: 19 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 27349176, 17217624).
Exposure time: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling UBE2C with ab252940 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in proliferative cells of mouse colon is observed (PMID: 15208666). The section was incubated with ab252940 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
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UBE2C was immunoprecipitated from 0.35 mg NIH:OVCAR-3 (Human ovary adenocarcinoma epithelial cell) whole cell lysate 10µg with ab252940 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab252940 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: NIH:OVCAR-3 whole cell lysate 10µg.
Lane 2: ab252940 IP in NIH:OVCAR-3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252940 in NIH:OVCAR-3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma epithelial cell) cells labeling UBE2C with ab252940 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in partial SW480 cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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: ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling UBE2C with ab252940 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in proliferative cells of rat colon is observed (PMID: 15208666). The section was incubated with ab252940 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
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All lanes : Anti-UBE2C antibody [EPR23165-31] (ab252940) at 1/1000 dilution
Lane 1 : T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate
Lane 2 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 20 kDa
Observed band size: 19 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 27349176, 17217624).
Exposure time: 3 minutes.
-
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling UBE2C with ab252940 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasm and nuclear staining in proliferative cells of human colon is observed (PMID: 15208666). The section was incubated with ab252940 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
-
UBE2C was immunoprecipitated from 0.35 mg SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10µg with ab252940 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab252940 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: SW480 whole cell lysate 10µg.
Lane 2: ab252940 IP in SW480 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252940 in SW480 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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