Key features and details
- Mouse monoclonal [BA-17] to Cytokeratin 19
- Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
製品名Anti-Cytokeratin 19 antibody [BA-17]
Cytokeratin 19 一次抗体 製品一覧
製品の詳細Mouse monoclonal [BA-17] to Cytokeratin 19
特異性Cytokeratin peptide 19 (40 kDa) in human tissue.
アプリケーション適用あり: Flow Cyt, ICC/IF, WB, IHC-P, IPmore details
- ICC/IF KO: MCF7 cells (MCF7-KRT19 KO used as a negative cell line).
保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.097% Sodium azide
Concentration information loading...
精製度Protein A purified
Our Abpromise guarantee covers the use of ab7755 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1-2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration.|
機能Involved in the organization of myofibers. Together with KRT8, helps to link the contractile apparatus to dystrophin at the costameres of striated muscle.
組織特異性Expressed in a defined zone of basal keratinocytes in the deep outer root sheath of hair follicles. Also observed in sweat gland and mammary gland ductal and secretory cells, bile ducts, gastrointestinal tract, bladder urothelium, oral epithelia, esophagus, ectocervical epithelium (at protein level). Expressed in epidermal basal cells, in nipple epidermis and a defined region of the hair follicle. Also seen in a subset of vascular wall cells in both the veins and artery of human umbilical cord, and in umbilical cord vascular smooth muscle. Observed in muscle fibers accumulating in the costameres of myoplasm at the sarcolemma in structures that contain dystrophin and spectrin.
配列類似性Belongs to the intermediate filament family.
発生段階Present in hair follicles at all stages of development.
ドメインThis keratin differs from all other IF proteins in lacking the C-terminal tail domain.
- Information by UniProt
- 40 kDa keratin intermediate filament antibody
- CK 19 antibody
- CK-19 antibody
ab7755 staining Cytokeratin 19 in wild-type MCF7 cells (top panel) and KRT19 knockout MCF7 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab7755 at 5µg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Human normal skin. Staining is observed in the cytoplasm (epidermal basal cells). Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
All lanes : Anti-Cytokeratin 19 antibody [BA-17] (ab7755) at 5 µg/ml
Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 2 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
Lane 3 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Observed band size: 44 kDa why is the actual band size different from the predicted?
Cytokeratin 19 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Mouse monoclonal to Cytokeratin 19 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7755.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 44kDa: Cytokeratin 19
ICC/IF image of ab7755 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7755, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Anti-Cytokeratin 19 antibody [BA-17] (ab7755) + Cell lysates prepared from human MCF-7 cells
Overlay histogram showing MCF7 cells stained with ab7755 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab7755, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab7755 は 16 報の論文で使用されています。
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