Name: Anti-Cytokeratin 18 antibody [C-04]
SKU: ab668
Rating: 5 (22 reviews)

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Flow Cytometry - Anti-Cytokeratin 18 antibody [C-04] (ab668)

Key features and details

Mouse monoclonal [C-04] to Cytokeratin 18
Suitable for: IHC-P, Flow Cyt
Reacts with: Human, Mammals
Isotype: IgG1

产品名称

Anti-Cytokeratin 18抗体[C-04]
参阅全部 Cytokeratin 18 一抗
描述

小鼠单克隆抗体[C-04] to Cytokeratin 18
宿主

Mouse
特异性

Human Cytokeratin
Tested Applications & Species

Application Species

Flow Cyt
Human

See all applications and species data
免疫原

Tissue, cells or virus corresponding to Cytokeratin 18. Cytoskeleton preparation of epidermal carcinoma cell line A431
阳性对照
- IHC-P: Human skin tissue. Flow Cyt: HCT 116 cells.

形式

Liquid
存放说明

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.
存储溶液

pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS
Concentration information loading...
纯度

Protein A purified
纯化说明

Purified from culture supernatant. Purity >95% by SDS-PAGE.
克隆

单克隆
克隆编号

C-04
同种型

IgG1
研究领域

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
Positive Controls
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab668 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用	Species
Flow Cyt	Human

应用	Ab评论	说明
IHC-P	(11)	Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt		Use a concentration of 1 - 4 µg/ml. Also see PMID 18946470. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

说明
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use a concentration of 1 - 4 µg/ml. Also see PMID 18946470. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

功能

Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection.
组织特异性

Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
疾病相关

Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].
序列相似性

Belongs to the intermediate filament family.
翻译后修饰

Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.
Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.
O-glycosylated at multiple sites; glycans consist of single N-acetylglucosamine residues.
细胞定位

Cytoplasm > perinuclear region.
Target information above from: UniProt accession P05783 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 3875 Human
- Omim: 148070 Human
- SwissProt: P05783 Human
- Unigene: 406013 Human
别名
- Cell proliferation inducing gene 46 protein antibody
- Cell proliferation inducing protein 46 antibody
- Cell proliferation-inducing gene 46 protein antibody
- CK 18 antibody
- CK-18 antibody
- CK18 antibody
- CYK 18 antibody
- CYK18 antibody
- Cytokeratin 18 antibody
- Cytokeratin endo B antibody
- Cytokeratin-18 antibody
- K 18 antibody
- K18 antibody
- K1C18_HUMAN antibody
- KA18 antibody
- Keratin 18 antibody
- Keratin 18, type I antibody
- Keratin D antibody
- keratin, type I cytoskeletal 18 antibody
- Keratin-18 antibody
- Krt18 antibody
see all

Flow Cytometry - Anti-Cytokeratin 18 antibody [C-04] (ab668)

Flow cytometric pattern (intracellular staining) of HeLa cells (Human epithelial cell line from cervix adenocarcinoma cell suspension) stained with ab668 (0.6 µg/ml concentration)
Immunohistochemistry paraffin embedded sections - Anti-Cytokeratin 18 antibody [C-04] (ab668)

Immunohistochemistry (paraffin-embedded sections) analysis of human kidney tissue labelling cytokeratin-18 with ab668 at 10µg/ml concentration.
Flow Cytometry - Anti-Cytokeratin 18 antibody [C-04] (ab668)

Flow cytometric analysis (intracellular staining) of HeLa cells (human epithelial cell line from cervix adenocarcinoma cell suspension) stained with 0.6 µg/ml ab668 (in red), and unstained HeLa cells (black).
Flow Cytometry - Anti-Cytokeratin 18 antibody [C-04] (ab668)

Overlay histogram showing HCT 116 (Human colorectal carcinoma cell line) cells stained with ab668 (red line).

The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab668, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 HCT 116 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [C-04] (ab668)This image is courtesy of an Abreview submitted by Carl Hobbs

ab668 staining Cytokeratin 18 in cat lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

Coloured arrowheads indicate positivity.
Good immunolabeling of bronchial tree epithelia (green), alveolar lining epithelium (red ).
Goblet cells are negative (asterisk).
Heavily stained structures are submucosal mucous glands.
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [C-04] (ab668)This image is courtesy of an Abreview submitted by Carl Hobbs

ab668 staining cytokeratin 18 in mouse epithelium tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

Composite image of d14 embryo shows developing epithelia.

Upper image: positive simple epithelium of renal tubules

Lower image: negative stratified epithelium of epidermis
See Abreview

Flow cytometry protocols
Immunohistochemistry protocols

Click here to view the general protocols

发表研究结果有使用 ab668？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab668 被引用在 99 文献中.

Zhao L et al. TDP-43 facilitates milk lipid secretion by post-transcriptional regulation of Btn1a1 and Xdh. Nat Commun 11:341 (2020). PubMed: 31953403
McCormick A et al. Placental endothelin-converting enzyme-1 is decreased in preeclampsia. Pregnancy Hypertens 20:108-110 (2020). PubMed: 32278308
Zhang B et al. Klf5 acetylation regulates luminal differentiation of basal progenitors in prostate development and regeneration. Nat Commun 11:997 (2020). PubMed: 32081850
Tamura R et al. XCL1 expression correlates with CD8-positive T cells infiltration and PD-L1 expression in squamous cell carcinoma arising from mature cystic teratoma of the ovary. Oncogene 39:3541-3554 (2020). PubMed: 32115573
Cheng L et al. Properties of an alginate-gelatin-based bioink and its potential impact on cell migration, proliferation, and differentiation. Int J Biol Macromol 135:1107-1113 (2019). PubMed: 31173833

View all Publications for this product

客户评价及客户问答

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提交评价提交问题

1-10 of 31 Abreviews or Q&A

Application

Immunocytochemistry

Sample

Human Cultured Cells (Endocervical epithelial cells)

Permeabilization

Yes - 0.75% Triton-X

Specification

Endocervical epithelial cells

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

提交于 Jan 28 2021

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (amnion epithelial cell)

Permeabilization

Specification

amnion epithelial cell

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

提交于 Feb 10 2021

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (Skin, oral epithelium)

Permeabilization

Specification

Skin, oral epithelium

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

提交于 Dec 29 2017

Application

Western blot

Sample

Human Cell lysate - whole cell (T24 cells)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

35 µg

Specification

T24 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

提交于 Jun 12 2017

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Sample

Pig Tissue sections (Small intestine)

Specification

Small intestine

Permeabilization

Fixative

Formaldehyde

Mr. Carl Hobbs

Verified customer

提交于 Sep 11 2014

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Sample

Cat Tissue sections (Lung)

Specification

Lung

Permeabilization

Fixative

Formaldehyde

Mr. Carl Hobbs

Verified customer

提交于 Jan 13 2014

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Sample

Goat Tissue sections (Lung)

Specification

Lung

Permeabilization

Fixative

Formaldehyde

Mr. Carl Hobbs

Verified customer

提交于 Jan 13 2014

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Blocking step

BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1:1000 · Temperature: RT°C

Sample

Rabbit Tissue sections (eye)

Specification

eye

Permeabilization

Fixative

Formaldehyde

Dr. Alex Zagariya

Verified customer

提交于 Jan 06 2014

Question

Thank you for your reply! I know Ab668 is a popular one, but most mouse
antibodies have background when stain on mouse tissue.
The Ab668 also has high background in our mouse olfactory epithelium
tissue. Please check the attachment file. That's why I want to order a
rabbit anti-ck18.
Our protocol as follows:
1. olfactory epithelium tissue fixed in 4%PFA at 4C overnight
wash and equilibrated in sucrose/PBS, embeded in OCT.
2. Cryosections were prepared at 10 um. 4%PFA postfixed for 5min.
3. Washed in PBS and blocked in 10% Normal Goat serum or Donkey serum plus
1%BSA. 4C overnight.
4. Ms-antiCK18(Ab668) 1:100 in blocking buffer 4C 48 hours
5. Washed in PBS 4x and incubated with Gt-anti Ms Alex fluo 546 Dilution
1000, incubated 1.5hour at Room temperature. Then wash in PBS 4X.
According to the publications, CK18 stain the cells on the apical layer in
Olfactory epithelium. But Ab668 stained everywhere.

Abcam community

Verified customer

Asked on Jan 31 2013

Answer

Hello! Please find below our mouse on mouse staining tips. This may help you to improve your results. Additionally I would recommend you reduce the primary incubation to overnight, and incubate the antibodies in a solution of TBS plus 1% BSA.

We cannot vouch for the Millipore version of the clone but we have tested ab32118 and found that it does not react in mouse, and have placed this information on our datasheet to inform our customers.

Tips for eliminating background when staining mouse tissue with a mouse monoclonal

View a pdf version of this page
Staining of mouse tissue using mouse antibody is a complicated process as high levels of background are often observed. It is notoriously difficult to eliminate this background.
Much of the background is caused by secondary antibody binding to endogenous mouse IgG in the tissue being stained, and to Fc receptors on B cells, plasma cells and macrophages.
Abcam is unable to guarantee monoclonal mouse antibodies on mouse tissue (unless stated on the datasheet). However, there are a few tips to try and reduce this background should mouse on mouse staining be necessary:

1. Blocking of endogenous IgG

Prepare tissue sections as usual.
At the usual blocking step, block with serum (from same species as the secondary antibody) for 30 min at room temperature.
Wash 3 X 2 min with PBS Tween 20.
Incubate sections with an unconjugated AffiniPure Fab fragment Anti-Mouse IgG (H+L) for 1 hr at room temperature, or overnight 4°C . Try this blocking antibody at 0.1 mg/ml although the optimal concentration will need to be assessed by the end user.Proceed with antibody staining.

2. Blocking endogenous Fc receptors
There are kits available for this which use F(ab) monomeric secondary antibodies.

Other tips that can be used to decrease general background:

Incubate sections with 1% Triton (in PBS) at room temperature for 30 min to ‘clean’ the tissue.
Use TBS –Tween 20 as a washing buffer. This often gives less background than using PBS-Tween 20.

Abcam Scientific Support

回复于 Jan 31 2013

Question

1. The concentration of sheep serum used is 1:1000, diluted in PBS+1% TritonX. We block at room temperature for 90 minutes.

2. Antigen retrieval was performed at 96C for 25 minutes. We haven’t performed a time series, we just went off several methods which recommended 25 minutes for antigen retrieval. There was no difference between the two pHs. The pH 10 buffer we used was in fact a Tris based buffer, not a citrate based buffer.

3. We haven’t tested the secondary antibody with known primary antibodies which work as yet, as our lab has limited primary antibodies raised in mouse. We do however have a goat-anti-mouse fluorescent antibody which does work, and we will be trying this in the near future.

Thanks,

Abcam community

Verified customer

Asked on Oct 02 2012

Answer

Thank you for your reply.

I am sorry to hear that you have difficulties getting satisfactory results form this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimize the results from ab668.

1.) I suggest to decrease the amount of Triton X used in the buffers. 0.025% Triton X-100 is used in standard protocols. Triton X is a quite harsh detergent.

2.) Secondary anti pan IgG antibodies sometimes bind some isotypes better than others. I therefore suggest to try a different secondary antibody and use an IgG1 specific antibody.

Please let me know how you are getting on.

Abcam Scientific Support

回复于 Oct 02 2012

1-10 of 31 Abreviews or Q&A

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Key features and details

概述

产品名称

描述

宿主

特异性

Tested Applications & Species

免疫原

阳性对照

性能

形式

存放说明

存储溶液

纯度

纯化说明

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

疾病相关

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

文献 (99)

客户评价及客户问答

Immunocytochemistry abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunohistochemistry (Frozen sections) abreview for Anti-Cytokeratin 18 antibody [C-04]

Western blot abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Cytokeratin 18 antibody [C-04]

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Cytokeratin 18 antibody [C-04]