MA5-12596

antibody from Invitrogen Antibodies

Targeting: KRT5 EBS2, KRT5A

Provider product page for MA5-12596

Western blot

Antibody data

Antibody Data
Antigen structure
References [42]
Comments [0]
Validations
Immunocytochemistry [4]
Immunohistochemistry [3]
Flow cytometry [1]
Other assay [10]

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Product number: MA5-12596 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cytokeratin 5 Monoclonal Antibody (XM26)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-12596 targets Cytokeratin 5 in IHC (P) applications and shows reactivity with Human samples. The MA5-12596 immunogen is recombinant protein corresponding to C-terminal 103 aa of cytokeratin 5.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: XM26
Vial size: 500 µL
Concentration: Conc. Not Determined
Storage: 4°C

KRT5 protein structure - MA5-12596 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Cytokeratin 5 was performed using 70% confluent log phase A-431 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 5 (XM26) Mouse Monoclonal Antibody (Product # MA5-12596) at 1:250 dilution 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) at 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 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: Immunofluorescence analysis of Cytokeratin 5 was performed using 70% confluent log phase A-431 cells. The cells were fixed with ice-cold acetone at 4°C for 5 minutes and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cytokeratin 5 Monoclonal Antibody (XM26) (Product # MA5-12596) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), 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: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoskeletal 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: Immunofluorescence analysis of Cytokeratin 5 was performed using 70% confluent log phase A-431 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 5 (XM26) Mouse Monoclonal Antibody (Product # MA5-12596) at 1:250 dilution 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) at 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 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: Immunofluorescence analysis of Cytokeratin 5 was performed using 70% confluent log phase A-431 cells. The cells were fixed with ice-cold acetone at 4°C for 5 minutes and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cytokeratin 5 Monoclonal Antibody (XM26) (Product # MA5-12596) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175), (1:2000 dilution), 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: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoskeletal 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: Immunohistochemical analysis of KRT5 was performed using formalin-fixed paraffin-embedded human skin (malignant melanoma) tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - Low pH (10X) (Product # 00-4955-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with or without Cytokeratin 5 Monoclonal Antibody (XM26) (Product # MA5-12596) at 1:100 dilution in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. ReadyProbes™ Tissue Autofluorescence Quenching Kit (Product # R37630) was used to quench autofluorescence from the tissues. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human mesothelioma stained with Keratin 5 antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Cytokeratin 5 was performed on cryosections of human skin tissue. Tissues were blocked in 10% normal goat serum in 1X PBS containing 0.1% Triton X-100 (1X PBS-T) for 1 hour at room temperature (RT). The tissues were labeled with a Cytokeratin 5 monoclonal antibody (clone XM26, green, Product # MA5-12596) diluted 1:50 in 3% normal goat serum in 1X PBS-T for 1 hour at RT, followed by detection with a Goat anti-Mouse IgG1, Alexa Fluor 488 secondary antibody (Product # A-21121) diluted 1:2000 in 3% normal goat serum in 1X PBS-T for 1 hour at RT. Nuclei (blue) were stained with DAPI, included in ProLong Gold Anti-Fade Mountant (Product # P36931). Images were taken on an inverted microscope at 20X magnification. Data courtesy of Dr. Jiyoon Lee at Indiana University School of Medicine.

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIG 7 SARS-CoV-2 infects ciliated and goblet epithelial cells but not basal and club cells. Epithelial cells of the mock- and SARS-CoV-2-infected HAE-ALI B9-20 cultures at 4 dpi (MOI = 0.2) were dissociated from the Transwell insert and cytospun onto slides. The cells on the slides were fixed, permeabilized, and immunostained with anti-NP and together with anti-beta-tubulin IV (A), and anti-MUC5AC (B), anti-cytokeratin 5 (C), and anti-SCGB1A1 (D), respectively. Confocal images were taken at a magnification of x63. Nuclei were stained with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Delivery of CCL2-neutralizing antibodies did not significantly affect macrophage infiltration or tumor angiogenesis. (A) Primary tumor tissues were co-immunofluorescent stained for CK5 (green) and F4/80 (red). Secondary antibody controls overlaid with 4',6-diamidino-2-phenylindole are shown. (B) Primary tumor tissues were immunostained for vWF8. Magnified insert shows positive staining. Staining was quantified by Image J. Statistical analysis was performed using two-tailed Student's t test. Statistical significance was determined by P < .05. ns = not significant. Mean +- SEM is shown. Scale bar = 200 mum. Figure 5

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 3 Figure Treatment of TRb-expressing MDA-MB-468 cells with T 3 represses cytokeratin genes. (A) Gene Set Enrichment Analysis performed between T 3 -treated 468-EV and 468-TRbeta expression analysis revealed Gene Ontology Biological Processes or Hallmark gene sets involving differentially expressed cytokeratins genes (adjusted p value < 1e-7). (B) Heatmap of all expressed KRT genes in 468-EV or 468-TRbeta cells following Vehicle or 10 nM T 3 treatment. (C) Normalized gene expression bar plots of the cytokeratin genes KRT5 and KRT14 from RNA-seq data, n = 3 for each condition. (D) and (E) Protein expression of KRT5 and KRT14 were evaluated by western blot, compared with beta-Actin as a loading control. The blot presented is representative of three independent experiments. **** p < 0.0001. TRbeta, thyroid hormone receptor beta [Color figure can be viewed at wileyonlinelibrary.com ]

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Keratin 5 and fibulin-4 were found in the amyloid deposits of Bowen's carcinoma and lichen amyloidosis skin. (a) The skin from a Bowen's carcinoma (patient 1) was stained with anti-fibulin-4, anti-keratin 5, and anti-fibrillin-1 antibodies (the left column) together with thioflavin T. Bar = 50 mum. ( b ) Paraffin-embedded sections (patient 2, lichen amyloidosis) were stained with anti-fibulin-4 (red) and anti-keratin 5 (green, data not shown) and with anti-keratin 5 and thioflavin T. Bar = 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fibulin-4 and keratin 5 were found in the amyloid deposits of lichen amyloidosis (patient 3). ( a ) Consecutive frozen sections were stained with anti-keratin 5 or with anti-fibulin-4, and amyloid deposits were detected with thioflavin T. Each merged image is also shown. ( b ) The staining with anti-fibulin-4 in the same section as the second row of a was shown at lower magnification. This image also shows the localization of fibulin-4 in normal human skin. Fibulin-4 localizes in elastic fibers and in the surrounding blood vessels, not only in the papil1ary dermis but also in the reticular dermis. Bar = 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Prolactin suppression of BCL6 and subsequently CK5 is mediated by the Stat5 pathway (A) Immunoblots of BCL6, CK5, phosphorylated-Stat5 (p-Stat5), Stat5a, and GAPDH protein levels in extracts of T47D cultures exposed to adenovirus carrying either Stat5a or LacZ control for 24 h and subsequently treated with vehicle (Cntrl), Prolactin (PRL), beta-Estradiol (E2), or R5020 (Pg) for 48 h. (B) Immunoblots of BCL6, CK5, phosphorylated-Stat5 (p-Stat5), Stat5, Jak2, and GAPDH protein levels in extracts of T47D cultures treated with non-target shRNA (NTC) or Jak2 shRNA for 48h, followed by addition of vehicle (Cntrl), Prolactin (PRL), beta-Estradiol (E2), or R5020 (Pg) for 48 h. (C) Immunoblots of BCL6, phosphorylated-ERK1/2 (p-ERK1/2), ERK1/2, and GAPDH protein levels in extracts of T47D cultures pre-treated with Pg for 24 h followed by incubation with or without prolactin (PRL) in the presence or absence of Mek inhibitor (U0126) for 3 and 6 h. (D) Immunoblots of BCL6, phosphorylated-Akt (p-Akt), Akt, and GAPDH protein levels in extracts of T47D cultures pre-treated with Pg for 24 h followed by incubation with or without prolactin (PRL) in the presence or absence of PI3K inhibitor (Ly294002) for 3 and 6 h.