- MA5-14428 - Invitrogen Antibodies KRT8 antibody

Submitted references Pituispheres Contain Genetic Variants Characteristic to Pituitary Adenoma Tumor Tissue.

Peculis R, Mandrika I, Petrovska R, Dortane R, Megnis K, Nazarovs J, Balcere I, Stukens J, Konrade I, Pirags V, Klovins J, Rovite V
Frontiers in endocrinology 2020;11:313

Delineation of retroperitoneal metastatic lymph nodes in ovarian cancer with near-infrared fluorescence imaging.

Pu T, Xiong L, Liu Q, Zhang M, Cai Q, Liu H, Sood AK, Li G, Kang Y, Xu C
Oncology letters 2017 Sep;14(3):2869-2877

Casein kinase 1 is recruited to nuclear speckles by FAM83H and SON.

Kuga T, Kume H, Adachi J, Kawasaki N, Shimizu M, Hoshino I, Matsubara H, Saito Y, Nakayama Y, Tomonaga T
Scientific reports 2016 Sep 29;6:34472

FAM83H and casein kinase I regulate the organization of the keratin cytoskeleton and formation of desmosomes.

Kuga T, Sasaki M, Mikami T, Miake Y, Adachi J, Shimizu M, Saito Y, Koura M, Takeda Y, Matsuda J, Tomonaga T, Nakayama Y
Scientific reports 2016 May 25;6:26557

Ambient Light Promotes Selective Subcellular Proteotoxicity after Endogenous and Exogenous Porphyrinogenic Stress.

Maitra D, Elenbaas JS, Whitesall SE, Basrur V, D'Alecy LG, Omary MB
The Journal of biological chemistry 2015 Sep 25;290(39):23711-24

Multiple oncocytic cystadenoma with intraluminal crystalloids in parotid gland: case report.

Başak K, Kroğlu K
Medicine 2014 Dec;93(27):e246

A novel mechanism of keratin cytoskeleton organization through casein kinase Iα and FAM83H in colorectal cancer.

Kuga T, Kume H, Kawasaki N, Sato M, Adachi J, Shiromizu T, Hoshino I, Nishimori T, Matsubara H, Tomonaga T
Journal of cell science 2013 Oct 15;126(Pt 20):4721-31

Glucose and SIRT2 reciprocally mediate the regulation of keratin 8 by lysine acetylation.

Snider NT, Leonard JM, Kwan R, Griggs NW, Rui L, Omary MB
The Journal of cell biology 2013 Feb 4;200(3):241-7

Keratin K18 increases cystic fibrosis transmembrane conductance regulator (CFTR) surface expression by binding to its C-terminal hydrophobic patch.

Duan Y, Sun Y, Zhang F, Zhang WK, Wang D, Wang Y, Cao X, Hu W, Xie C, Cuppoletti J, Magin TM, Wang H, Wu Z, Li N, Huang P
The Journal of biological chemistry 2012 Nov 23;287(48):40547-59

Aging modulates susceptibility to mouse liver Mallory-Denk body formation.

Hanada S, Harada M, Abe M, Akiba J, Sakata M, Kwan R, Taniguchi E, Kawaguchi T, Koga H, Nagata E, Ueno T, Sata M
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 2012 Jun;60(6):475-83

Derivation and characterization of matched cell lines from primary and recurrent serous ovarian cancer.

Létourneau IJ, Quinn MC, Wang LL, Portelance L, Caceres KY, Cyr L, Delvoye N, Meunier L, de Ladurantaye M, Shen Z, Arcand SL, Tonin PN, Provencher DM, Mes-Masson AM
BMC cancer 2012 Aug 29;12:379

Keratin hypersumoylation alters filament dynamics and is a marker for human liver disease and keratin mutation.

Snider NT, Weerasinghe SV, Iñiguez-Lluhí JA, Herrmann H, Omary MB
The Journal of biological chemistry 2011 Jan 21;286(3):2273-84

Hedgehog overexpression leads to the formation of prostate cancer stem cells with metastatic property irrespective of androgen receptor expression in the mouse model.

Chang HH, Chen BY, Wu CY, Tsao ZJ, Chen YY, Chang CP, Yang CR, Lin DP
Journal of biomedical science 2011 Jan 18;18:6

Molecular characterization of breast cancer in young Brazilian women.

Carvalho LV, Pereira EM, Frappart L, Boniol M, Bernardo WM, Tarricone V, Tavtigian S, Southey MC
Revista da Associacao Medica Brasileira (1992) 2010 May-Jun;56(3):278-87

Molecular characterization of breast cancer in young Brazilian women.

Carvalho LV, Pereira EM, Frappart L, Boniol M, Bernardo WM, Tarricone V, Tavtigian S, Southey MC
Revista da Associacao Medica Brasileira (1992) 2010 May-Jun;56(3):278-87

Characterization of three new serous epithelial ovarian cancer cell lines.

Ouellet V, Zietarska M, Portelance L, Lafontaine J, Madore J, Puiffe ML, Arcand SL, Shen Z, Hébert J, Tonin PN, Provencher DM, Mes-Masson AM
BMC cancer 2008 May 28;8:152

Switch in Fas-activated death signaling pathway as result of keratin 8/18-intermediate filament loss.

Gilbert S, Ruel A, Loranger A, Marceau N
Apoptosis : an international journal on programmed cell death 2008 Dec;13(12):1479-93

Gene expression profiling of extracellular matrix as an effector of human hepatocyte phenotype in primary cell culture.

Page JL, Johnson MC, Olsavsky KM, Strom SC, Zarbl H, Omiecinski CJ
Toxicological sciences : an official journal of the Society of Toxicology 2007 Jun;97(2):384-97

Bispecific and human disease-related anti-keratin rabbit monoclonal antibodies.

Tao GZ, Nakamichi I, Ku NO, Wang J, Frolkis M, Gong X, Zhu W, Pytela R, Omary MB
Experimental cell research 2006 Feb 15;312(4):411-22

Vitamin K3 (menadione)-induced oncosis associated with keratin 8 phosphorylation and histone H3 arylation.

Scott GK, Atsriku C, Kaminker P, Held J, Gibson B, Baldwin MA, Benz CC
Molecular pharmacology 2005 Sep;68(3):606-15

Keratins modulate c-Flip/extracellular signal-regulated kinase 1 and 2 antiapoptotic signaling in simple epithelial cells.

Gilbert S, Loranger A, Marceau N
Molecular and cellular biology 2004 Aug;24(16):7072-81

Functional testing of keratin 14 mutant proteins associated with the three major subtypes of epidermolysis bullosa simplex.

Sørensen CB, Andresen BS, Jensen UB, Jensen TG, Jensen PK, Gregersen N, Bolund L
Experimental dermatology 2003 Aug;12(4):472-9

Functional testing of keratin 14 mutant proteins associated with the three major subtypes of epidermolysis bullosa simplex.

Sørensen CB, Andresen BS, Jensen UB, Jensen TG, Jensen PK, Gregersen N, Bolund L
Experimental dermatology 2003 Aug;12(4):472-9

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1) and A-431 (Lane 2).The blot was probed with Anti-Cytokeratin 8 Mouse monoclonal Antibody (Product # MA5-14428, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 52 kDa band corresponding to Cytokeratin 8 was observed in cell lines tested. An additional band at 48 kDa band corresponding to other Cytokeratin was also observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of KRT8 was achieved by transfecting Caco-2 with KRT8 specific siRNAs (Silencer® select Product # S7969, S7970). Western blot analysis (Fig. a) was performed using Whole cell extracts from the KRT8 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cytokeratin 8 Monoclonal Antibody (TS1) (Product # MA5-14428, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to KRT8.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Cytokeratin 8 Monoclonal Antibody (TS1) (Product # MA5-14428) and a 53 kDa band corresponding to KRT8 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), MCF7 (Lane 3), HaCaT (Lane 4) and Caco-2 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cytokeratin 8 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin 8 (TS1) Mouse Monoclonal Antibody (Product # MA5-14428) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Cytokeratin 8 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR745510_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Immunofluorescence analysis was performed on wild type HeLa cells (panel a,d), HeLa Cas9 control cells (panel b,e) and HeLa Cytokeratin 8KO cells (panel c, f). Cells were fixed, permeabilized, and labelled with Cytokeratin 8 Monoclonal Antibody (TS1) (SP4) (Product # MA5-144288203, 1:100 dilution), followed by Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 488 (Product # A32723, 1:2,000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal (panel c,f) upon CRISPR mediated knockout (KO) confirms that antibody is specific to Cytokeratin 8 (green). The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of KRT8 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Cytokeratin 8 Monoclonal Antibody (TS1) (Product # MA5-14428) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723, 1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Blue) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Intermediate filaments localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human breast carcinoma stained with Keratin 8 antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Cytokeratin 8 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Cytokeratin 8 Mouse Monoclonal Antibody (MA5-14428, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. Staining of imaging-positive lymph nodes. H&E staining of imaging-positive lymph nodes at (A) x100 and (B) x400 magnification. IHC analysis of human CK8 in imaging-positive lymph nodes at (C) x100 and (D) x400 magnification. IHC analysis of human CK7 in imaging-positive lymph nodes at (E) x100 and (F) x400 magnification. Positive staining of tumor cells was sporadically distributed. H&E, hematoxylin and eosin; IHC, immunohistochemistry; CK, cytokeratin.

MA5-14428

Antibody data