MA5-11344

antibody from Invitrogen Antibodies

Targeting: KRT15 CK15, K15, K1CO

Provider product page for MA5-11344

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [51]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [3]
Immunohistochemistry [3]
Other assay [12]

Submit

Product number: MA5-11344 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cytokeratin 15 Monoclonal Antibody (LHK15)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA5-11344 targets Cytokeratin 15 in ICC/IF, IHC-P and Western blot applications and shows reactivity with Bovine, Human, mouse, Non-human primate and Rat samples. The MA5-11344 immunogen is a 17-mer synthetic peptide from the C-terminus of human cytokeratin 15.
Reactivity: Human, Mouse, Rat, Bovine
Host: Mouse
Isotype: IgG
Antibody clone number: LHK15
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

KRT15 protein structure - MA5-11344 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cytokeratin 15 (green) showing staining in the cytoplasm of COS7 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cytokeratin 15 (green) showing staining in the cytoplasm of Hela cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Cytokeratin 15 (green) showing staining in the cytoplasm of HepG2 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytokeratin 15 showing staining in the cytoplasm of paraffin-treated human prostate tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytokeratin 15 showing staining in the cytoplasm of paraffin-treated human skin tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytokeratin 15 showing staining in the cytoplasm of paraffin-treated mouse skin tissue (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Cytokeratin 15 monoclonal antibody (Product # MA5-11344) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Localisation of stem cells in human nails. Side view, components of the human nail a Red circles are cells in nail proximal folds (NPFs); green circles are cells in the nail matrix (Mx). We found K15 + label-retaining cells in human nails in both the NPF and Mx. The diagrammatic drawing shows the tangent plane of the slice. b H&E staining of human nail side facing showing the nail bed (NB), nail root at the Mx and the NPF junction (scale bar = 1000 mum). c, d Immunofluorescence staining for K15 (green) and Ki67 (red); cell nuclei were stained blue by DAPI, image of separately staining of Ki67 in the upper left corner. The positive expression of K15 extended from the PF to the proximal Mx (scale bar = 200 mum). e Immumohistochemical staining for K15 (brown) (scale bar = 1000 mum). f Magnified photographs of immumohistochemical staining for K15 (brown) (scale bar = 200 mum). (Color figure online)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Expression of stem cell markers and the difference between young and aged participants in human nail stem cells. a Immunohistochemical staining for K14, K15, and K19 in human nails. These three markers were all expressed in the NPF and Mx in both groups. Compared to K14 and K15, the expression of K19 was relatively weak, but there was no significant difference between the two groups (scale bar = 400 mum). b Immunohistochemical staining for CD29, CD34, and Lgr6 in human nails. The markers were expressed in the same area (scale bar = 400 mum). c Proportion of positive cells stained for K14, K15, K19, CD29, CD34, and Lgr6 in analysed slices (n = 58, p > 0.05), illustrating no significant difference between the two groups. d Diagrammatic drawing of the nail root. The area in the blue and black box was used to determine the percentage of positive cells. e Co-immunofluorescence staining for K14 and K15 (scale bar = 400 mum). (Color figure online)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Comparison of cell proliferation between the young and aged groups. a Comparison of Ki67-positive cells with K15 positivity in the two groups (scale bar = 200 mum), image of separately staining of Ki67 in the upper left corner. c Ki67-positive cell count results; the young group was higher than the aged group (n = 58, p = 0.0005). b, d Comparison of Ki67-positive cells with K14 positivity in the two groups and Ki67-positive cell count results (scale bar = 200 mum), image of separately staining of Ki67 in the upper left corner. Similar results were obtained for both K14 and K15 staining (n = 58, p = 0.002). **P < 0.01 ***P < 0.001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 Comparison of nail stem cell proliferation and viability between the young and aged groups. a - h, i Nail stem cell characterisation. When cultured in DKSFM, the cells tended to clump together ( a, b ; scale bar = 1000 mum) and showed high proliferation capacity ( c, d, i ; scale bar = 200 mum). e - h Immunofluorescence staining for K14, K15 (green), and cell nuclei (DAPI/blue) (scale bar = 200 mum). j After seeding, cell viability was evaluated by CCK assays; the cell metabolic activity of the young group was significantly higher than the aged group (p = 0.04). k - m Flow cytometric analysis of total (left side graphs) or K15(+) (right side graphs) cell cycle status in young versus old nail preparations (p < 0.05). *P < 0.05. (Color figure online)