Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 02, 2021

Anti-UFC1 antibody [EPR15014-102] (ab189252)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR15014-102] to UFC1
Suitable for: IP, ICC/IF, WB, IHC-P
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-UFC1 antibody [EPR15014-102]
See all UFC1 primary antibodies
Description

Rabbit monoclonal [EPR15014-102] to UFC1
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal liver tissue lysate and HEK-293T, HAP1, U87-MG, MCF7, A549, RAW 264.7, and NIH 3T3 cell lysates. IHC-P: Human colon and human transitional cell carcinoma of bladder tissues. ICC/IF: MCF7 cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR15014-102
Isotype

IgG
Research areas

Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)

All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : UFC1 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 19 kDa
Observed band size: 20 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab189252 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab189252 was shown to react with UFC1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266814 (knockout cell lysate ab257781) was used. Wild-type HEK-293T and UFC1 HEK-293T KO cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab189252 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon tissue labeling UFC1 with ab189252 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)

All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/10000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : UFC1 knockout HAP1 cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 19 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab189252 observed at 20 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab189252 was shown to react with UFC1 in wild-type HAP1 cells in western blot. Loss of signal was observed when UFC1 knockout sample was used. Wild-type HAP1 and UFC1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab189252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)

All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/20000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : U87-MG cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : A549 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 19 kDa
Western blot - Anti-UFC1 antibody [EPR15014-102] (ab189252)

All lanes : Anti-UFC1 antibody [EPR15014-102] (ab189252) at 1/5000 dilution

Lane 1 : RAW 264.7 cell lysate
Lane 2 : NIH 3T3 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 19 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFC1 antibody [EPR15014-102] (ab189252)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling UFC1 with ab189252 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-UFC1 antibody [EPR15014-102] (ab189252)

Immunofluorescent analysis of MCF7 cells(4% paraformaldehyde-fixed) labeling UFC1 with ab189252 at 1/100 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).
Immunoprecipitation - Anti-UFC1 antibody [EPR15014-102] (ab189252)

Western blot analysis of immunoprecipitation pellet from MCF7cell lysate immunoprecipitated using ab189252 at 1/50 dilution (lane 1) or PBS control (lane 2).
Secondary: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.
Anti-UFC1 antibody [EPR15014-102] (ab189252)

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