Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR21084] to Vimentin (phospho S56)
Suitable for: WB, Dot blot, ICC/IF, IP, Flow Cyt
Reacts with: Human

Product name

Anti-Vimentin (phospho S56) antibody [EPR21084]
See all Vimentin primary antibodies
Description

Rabbit monoclonal [EPR21084] to Vimentin (phospho S56)
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate. ICC/IF: HeLa cells. IP: HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate.
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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21084
Isotype

IgG
Research areas

Western blot - Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

All lanes : Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673) at 1/1000 dilution

Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 54 kDa
Observed band size: 57 kDa
why is the actual band size different from the predicted?

Exposure time: 23 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Flow Cytometry - Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

Ab217673 staining Vimentin (phospho S56) in HeLa (human cervix adenocarcinoma epithelial cell) cells by Flow Cytometry. Cells were fixed using 80% Methanol and permeabilized with 0.1% Tween-20. The sample was incubated with primary antibody at 1:500 dilution. An Alexa Fluor^®488 Goat anti-rabbit IgG (ab150077) was used as a secondary antibody. Rabbit monoclonal IgG (ab172730) was used as an isotype control (left). Cells were pre-treated with 20 μg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and PI (Propidium iodide). Vimentiin (phospho S56) is highly expressed in mitotic cells. (PMID: 16260496)
Dot Blot - Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

Dot blot analysis of Vimentin (phospho S56) labeled with ab217673 at 1/1000 dilution.

Lane 1: Vimentin (phospho S56) peptide.
Lane 2: Vimentin non-phospho peptide.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 1 minute.

The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
Immunocytochemistry/ Immunofluorescence - Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Vimentin (phospho S56) with ab217673 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing strong positive staining in HeLa cells in M phase.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunoprecipitation - Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

Vimentin (phospho S56) was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate with ab217673 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217673 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate 10 μg (Input).
Lane 2: ab217673 IP in HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217673 in HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)

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