Anti-Cytokeratin 13抗体[AE8]
参阅全部 Cytokeratin 13 一抗
小鼠单克隆抗体[AE8] to Cytokeratin 13
Mouse
适用于: IHC-Fr, ICC/IF, WB, IHC-Pmore details
与反应: Human
预测可用于: Mouse, Rabbit
Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.
WB: A431 whole cell lysate. ICC/IF: A431 cells. IHC-P: Human tonsil. IHC-Fr: Human tonsil.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Protein G purified
This antibody is specific for Cytokeratin 13, which is a marker for oesophageal type differentiation which is expressed by various internal stratified epithelia.
单克隆
AE8
P3-X63 Ag8.3
IgG
kappa
Abpromise™承诺保证使用ab16112于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
IHC-Fr | Use a concentration of 1 µg/ml. | |
ICC/IF | (1) | Use a concentration of 1 - 5 µg/ml. PubMed: 25076852 |
WB | Use a concentration of 1 µg/ml. Predicted molecular weight: 50 kDa. | |
IHC-P | (3) | Use a concentration of 0.05 µg/ml. |
Entrez Gene: 3860 Human
Entrez Gene: 16663 Mouse
Omim: 148065 Human
SwissProt: P13646 Human
SwissProt: P08730 Mouse
Unigene: 654550 Human
Unigene: 4646 Mouse
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Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
All lanes : Anti-Cytokeratin 13 antibody [AE8] (ab16112) at 1 µg/ml
Lane 1 : Wild-type A431 cell lysate
Lane 2 : KRT13 knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 51 kDawhy is the actual band size different from the predicted?
False colour image of Western blot: Anti-Cytokeratin 13 antibody [AE8] staining at 1 µg/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab16112 was shown to bind specifically to Cytokeratin 13. A band was observed at 51 kDa in wild-type A431 cell lysates with no signal observed at this size in Krt13 knockout cell line ab269483 (knockout cell lysate ab269647). To generate this image, wild-type and Krt13 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
IHC image of Cytokeratin 13 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16112, 0.05 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
IHC image of Cytokeratin 13 staining in a section of frozen normal human tonsil*. The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab16112 at 1µg/ml. The section was then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488), 1/1000)) (shown in green) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. The secondary-only control insert image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
Western blot - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
Anti-Cytokeratin 13 antibody [AE8] (ab16112) at 1 µg/ml + A431 whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 50 kDa
This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab16112 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 13 antibody [AE8] (ab16112)
ab16112 staining Cytokeratin 13 in A431 cells. The cells were fixed with Methanol (5min), permeabilized with 0.1%PBS-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 overnight at +4°C with ab16112 at 5μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 13 antibody [AE8] (ab16112)This image is courtesy of an anonymous Abreview
ab16112 staining Cytokeratin 13 in Human pharynx tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (undiluted) for 1 hour at 20°C. An undiluted HRP-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody.