Ab52635

Manufactured by Abcam

Sourced in United Kingdom, United States

Ab52635 is a primary antibody that targets a specific protein. It is designed for use in various laboratory applications.

Automatically generated - may contain errors

Lab products found in correlation

Ab76318, by Abcam (7 mentions) Ab7800, by Abcam (4 mentions) Ab238132, by Abcam (3 mentions) Ab283654, by Abcam (3 mentions) Gb121141, by Proteintech (3 mentions) Sc 53876, by Santa Cruz Biotechnology (3 mentions) Ab53280, by Abcam (3 mentions) Ab15580, by Abcam (3 mentions) Ab185628, by Abcam (2 mentions) Ab201015, by Abcam (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Pvdf membrane, by GE Healthcare (2 mentions) Lsm 700, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Ab93652, by Abcam (1 mentions) Ab109320, by Abcam (1 mentions) Ab13680, by Abcam (1 mentions) Ecl substrate, by Merck Group (1 mentions) Horseradish peroxidase hrp conjugated anti rabbit igg, by Santa Cruz Biotechnology (1 mentions) Inhibitor cocktail, by Selleck Chemicals (1 mentions)

37 protocols using ab52635

Immunohistochemical Analysis of Bladder Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Paraffin sections were cut (5-mm thick), dewaxed in xylene, and treated with an ethanol gradient. The sections were stained with hematoxylin and eosin or with antibodies against keratin 5 (Krt5) (Ab52635, 1:100; Abcam, Cambridge, UK), keratin 14 (Krt14) (Ab7800, 1:200; Abcam), forkhead box A1 (Foxa1) (Ab170933, 1:200; Abcam), and epidermal growth factor receptor (Ab52894, 1:200; Abcam).
For immunofluorescence, antigen retrieval was performed with citrate buffer (pH 6) before primary antibody incubation at 4 C overnight. Sections were stained with primary antibodies against Upk (Ab78196, 1:200; Abcam), Krt5 (Ab52635, 1:100; Abcam), and RFP (Ab62341, 1:50; Abcam), and secondary antibodies Alexa Fluore488 and Alexa Fluore594 (10 mg/mL; Thermo Fisher Scientific, Paisley, UK) for 1 hour at room temperature. Slides were mounted with Mowiol 488 (Sigma-Aldrich, Dorset, UK) containing DAPI (Vectashield Mounting Medium with DAPI, H-1200; Vector Laboratories, Burlingame, CA). Images were taken on a Leica confocal laser microscope (TCS SP8; Leica Microsystems, Wetzler, Germany).
Histopathologic evaluation of the bladder tissues was performed in a blind fashion by one pathologist at Kyoto Histopathology Study Group, Inc., and by two trained urologists (N.M. and T.K.), based on the 2004 World Health Organization Classification of Bladder Tumors.

Masuda N., Murakami K., Kita Y., Hamada A., Kamada M., Teramoto Y., Sakatani T., Matsumoto K., Sano T., Saito R., Okuno Y., Ogawa O, & Kobayashi T. (2020). Trp53 Mutation in Keratin 5 (Krt5)-Expressing Basal Cells Facilitates the Development of Basal Squamous-Like Invasive Bladder Cancer in the Chemical Carcinogenesis of Mouse Bladder. The American journal of pathology, 190(8).

+ Open protocol

+ Expand

Protein Expression Analysis in Skin

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The protein levels of KRT5, KRT14, KRT1, KRT10, STAT1, and S100A7 were examined by immunoblotting following the methods described before^{34 (link)} with primary antibodies against KRT5 (ab52635; Abcam, Cambridge, MA, USA), KRT14 (ab7800, Abcam), KRT1 (ab93652, Abcam), KRT10 (ab76318, Abcam), STAT1 (ab109320, Abcam), and S100A7 (ab13680, Abcam) and the HRP-conjugated secondary antibody. Signals were visualized using enhanced chemiluminescent (ECL) substrates (Millipore, MA, USA) with normalization to GAPDH.

Luo M., Huang P., Pan Y., Zhu Z., Zhou R., Yang Z, & Wang C. (2021). Weighted gene coexpression network and experimental analyses identify lncRNA SPRR2C as a regulator of the IL-22-stimulated HaCaT cell phenotype through the miR-330/STAT1/S100A7 axis. Cell Death & Disease, 12(1), 86.

+ Open protocol

+ Expand

Wound Regeneration and Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The samples were fixed with 4% paraformaldehyde at least 48 hours before ethanol and xylene dehydration. H&E staining and Masson’s trichrome staining were performed for the observation of re-epithelialization and collagen fiber deposition. Immunofluorescence staining for cytokeratin 5 (ab52635, Abcam, 1:200), cytokeratin 10 (ab76318, Abcam, 1:150), cytokeratin 17 (17516-1-AP, Proteintech, 1:200), TWIST2 (66544-1-Ig, Proteintech, 1:100), Ki67 (Servicebio, GB121141, 1:100), SCD1 (28678-1-AP, Proteintech, 1:200), CRABP1 (13163 S, Cell Signaling, 1:100), MEST (11118-1-AP, Proteintech, 1:100) were performed for to assess wound regeneration. For the evaluation the infiltration of immune cells, immunohistochemistry staining for CD3 (14-0032-82, Thermo Fisher Scientific, 1:100), CD68 (ab283654, Abcam, 1:100), Ly6G (ab238132, Abcam, 1:2000) and immunofluorescent staining for F4/80 (29414-1-AP, Proteintech, 1:200), CD206 (360017, Zenbio, 1:100), FOXP3 (sc-53876, Santa Cruz Biotechnology, 1:100), were performed.

Yang Y., Chu C., Liu L., Wang C., Hu C., Rung S., Man Y, & Qu Y. (2023). Tracing immune cells around biomaterials with spatial anchors during large-scale wound regeneration. Nature Communications, 14, 5995.

+ Open protocol

+ Expand

Histological and Immunohistochemical Analysis of Mammary Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mammary glands and tumors were fixed overnight at 4 °C with 4% PFA in PBS. For histological analysis, the tissues were embedded in paraffin and 5 μm tissue sections were obtained. Tissue sections were stained with hematoxylin and eosin (H/E) and histology images were acquired using a Nikon E800 upright widefield/fluorescence microscope. For immunohistochemical staining (IHC), antigen retrieval was first performed by heating for 5 min at 120 °C in a sodium citrate buffer at pH 6, followed by blocking for 1 h at RT with 3% BSA in PBS and staining overnight at 4 °C with anti-Cytokeratin 8 at 1:250 (rabbit, Abcam, Cambridge, UK, ab53280) and anti-Cytokeratin 5 at 1:250 (rabbit, Abcam, ab52635). Endogenous peroxidase activity was inhibited by incubating with 3% hydrogen peroxide for 10 min. In a final step, sections were incubated with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Santa Cruz Biotechnology, Dallas, TX, USA) for 1 h at RT, washed and treated with the substrate 3,3’-Diaminobenzidine (DAB) for 2–5 min in the dark. Hematoxylin was used for counterstaining.

Kolokotroni A., Gkikopoulou E., Rinotas V., Ntari L., Zareifi D., Rouchota M., Sarpaki S., Lymperopoulos I., Alexopoulos L.G., Loudos G., Denis M.C., Karagianni N, & Douni E. (2023). Α Humanized RANKL Transgenic Mouse Model of Progestin-Induced Mammary Carcinogenesis for Evaluation of Novel Therapeutics. Cancers, 15(15), 4006.

+ Open protocol

+ Expand

Airway Organoid Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The airway organoids were lysed in ice-cold RIPA buffer (Beyotime, Shanghai, CA, P0013C) containing an inhibitor cocktail (Bimake, Houston, TX, USA, B14001) with ultrasonic waves and incubated on ice for 40 min. Next, the samples were added to a 5× sodium dodecyl sulfate (SDS) loading buffer and heated at 95 °C. The denatured proteins were separated on 3–8% Tris-acetate gel (Thermo Fisher, Waltham, MA, USA, EA0375BOX) and transferred to a 0.45-µm pore size polyvinylidene difluoride (PVDF) membrane (Millipore) at 40 V overnight. Subsequently, the membranes were blocked in 5% skimmed milk for 1 h at RT and incubated with primary antibodies, GAPDH (adcam, ab181602), DNAH5 (adcam, ab234826) and Krt5 (abcam, ab52635) overnight at 4 °C. The following day, the samples were washed and incubated with a goat anti-mouse IgG secondary antibody-HRP (1:5000, Thermo Fisher, 32230) and goat anti-rabbit IgG secondary antibody-HRP (1:5000, Thermo Fisher, 6120) in 5% skimmed milk at RT for 1 h. An ECL chemical substrate (Millipore) was used to visualize the developed immunoblot.

Yang W., Chen L., Guo J., Shi F., Yang Q., Xie L., Lu D., Li Y., Luo J., Wang L., Qiu L., Chen T., Li Y., Zhang R., Chen L., Xu W, & Liu H. (2022). Multiomics Analysis of a DNAH5-Mutated PCD Organoid Model Revealed the Key Role of the TGF-β/BMP and Notch Pathways in Epithelial Differentiation and the Immune Response in DNAH5-Mutated Patients. Cells, 11(24), 4013.

+ Open protocol

+ Expand

Quantifying Cytokeratin Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The sections were pretreated with 1% BSA in PBS containing 0.1% Triton X-100 for 1 hour, incubated in 1% Tween 20 for 20 min, and washed again with PBS. The sections were subsequently analyzed for Krt10 and Krt5, according to the manufacturers’ instructions. Sections were briefly incubated for 30 min in the dark, and excessive dye was rinsed off using PBS. Sections were then incubated with the antibody isotypes to exclude false-positive staining. Double immunofluorescence staining with primary antibodies against cytokeratin 10 (ab76318, Abcam, 1:150) and cytokeratin 5 (ab52635, Abcam, 1:200) and with secondary antibodies (GB25303 and GB21303, 1:400; Servicebio, Wuhan, China) was performed. The immunostained specimens were further subjected to Hoechst 33258 staining (G1011, Servicebio). At least three parallel sections were observed using ortho-fluorescent microscopy and imaging system (Nikon, Tokyo, Japan). Fluorescence area measurements were conducted at five random sites of regenerated epithelia using CaseViewer 2.1 and Image-Pro Plus 7.0 (n = 5).

Hu C., Chu C., Liu L., Wang C., Jin S., Yang R., Rung S., Li J., Qu Y, & Man Y. (2021). Dissecting the microenvironment around biosynthetic scaffolds in murine skin wound healing. Science Advances, 7(22), eabf0787.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Hydrogels and Decellularized Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hydrogels and decellularized tissues were fixed in 10% neutral buffered formalin for 1 hr at room temperature, dehydrated, paraffin embedded and cut into 5μm sections. For immunostaining, sections were deparrafinized, rehydrated and subjected to heat mediated antigen retrieval in pH 9.0 Tris-EDTA buffer containing 0.05% v/v Tween 20 for 20 mins using a steam cooker. The following antibodies were used: rabbit polyclonal anti-ki67 (3.5μg/ml; Abcam; ab15580) and rabbit monoclonal (EP1601Y) anti-cytokeratin 5 (1.22μg/ml; Abcam; ab52635), and polyclonal goat anti-rabbit alexafluor 488 conjugated secondary (1μg/ml; ThermoFisher; A32731. Sections were all counterstained with DAPI. All images were taken on a Zeiss Axio Observer Z1 microscope and analyzed using the Zeiss Zen software. Cell counts were performed manually and fluorescence intensity was measured using the Zen software on a minimum of 6 random locations per field of view across a minimum of 4 sections of 3 separate samples. For Laminin 111/112 measurements, hydrogels were formed in a chamber slide, and stained with a rabbit polyclonal anti laminin 111/112 antibody (10μm/ml; Abcam; ab7463) and a goat alexafluor 488 conjugated secondary as described above. Fluorescence intensity was performed as described above across 3 independent samples.

Mollica P.A., Creech E., Reid J.A., Zamponi M., Sullivan S.M., Palmer X.L., Sachs P.C, & Bruno R.D. (2019). 3D bioprinted mammary organoids and tumoroids in human mammary derived ECM hydrogels. Acta biomaterialia, 95, 201-213.

+ Open protocol

+ Expand

Protein Isolation and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein isolation from whole-cell lysates was performed by using cell lysis buffer (Cell Signaling Technology, Danvers, MA) with 0.5% protease inhibitor cocktail (P8340, Sigma-Aldrich), 1% phosphatase inhibitor cocktail 2 (P5726, Sigma-Aldrich), and 1% phosphatase inhibitor cocktail 3 (P004, Sigma-Aldrich) on ice for 30 min. Protein extracts were separated by 7.5% SDS-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA). The membranes were blocked with 5% nonfat milk and immunoblotted with primary antibody of p63 (20697-1-AP, Proteintech, Wuhan, China), OCT4 (11263-1-AP, Proteintech), keratin 5 (K5, Ab52635, Abcam, US), keratin 14 (K14, 10143-1-AP, Proteintech), SHH (20697-1-AP, Proteintech), NRF2 (sc-13032, Santa Cruz Biotechnology, Santa Cruz, CA), or β-actin (sc-69879, Santa Cruz Biotechnology) overnight at 4°C and with secondary antibodies (Santa Cruz Biotechnology) for 1 h at RT. Protein bands were detected with the Chemistar ECL Western Blotting Substrate (180-5001, Tanon, Shanghai, China), and the image was captured by 5500 Automatic Chemiluminescence Imaging System (Tanon).

Wu X., Yang B., Hu Y., Sun R., Wang H., Fu J., Hou Y., Pi J, & Xu Y. (2017). NRF2 Is a Potential Modulator of Hyperresistance to Arsenic Toxicity in Stem-Like Keratinocytes. Oxidative Medicine and Cellular Longevity, 2017, 7417694.

+ Open protocol

+ Expand

Immunostaining Antibody Panel for Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

CAV1 BD6100407 (clone 2297) (1/1000 for IHC, 1/300 fluorescence). ITGB1 (EP1041Y; Ab52971; 1/500 IHC, 1/100 fluorescence). E-cadherin from CST (24E10, 1/300 fluorescence). The integrin antibodies ITGA2 (Novus, NBP1-96715, 1/200) and ITGA6 (Novus, NBP1-85747, 1/200). The Pan-CK antibody (C-11, Abcam, ab7753, 1 to 1500) and the KRT5 antibody (Abcam, ab52635, 1 to 2000).

Pellinen T., Blom S., Sánchez S., Välimäki K., Mpindi J.P., Azegrouz H., Strippoli R., Nieto R., Vitón M., Palacios I., Turkki R., Wang Y., Sánchez-Alvarez M., Nordling S., Bützow A., Mirtti T., Rannikko A., Montoya M.C., Kallioniemi O, & del Pozo M.A. (2018). ITGB1-dependent upregulation of Caveolin-1 switches TGFβ signalling from tumour-suppressive to oncogenic in prostate cancer. Scientific Reports, 8, 2338.

+ Open protocol

+ Expand

Organoid Characterization Using IHC Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cytoblocks were prepared from the pellets of organoids by adding plasma and thrombin in order to obtain a solid matrix. Once solidified, the organoids were fixed in 10% formalin (Thermo Scientific, 5701) and embedded in paraffin as normal tissue. Sections of 4 μm were used for IHC analyses and hematoxylin and eosin (H&E) staining (Diapath, C0303) and (Diapath, C0363), respectively. Once dried, the sections were treated with OTTIX plus solution (Diapath, X0076) and OTTIX shaper solution (Diapath, X0096) to dewax and rehydrate the sections. Antigen retrieval was performed using pH 6 solutions at 98 °C for 20 min. Next, the endogenous peroxidases and non-specific-binding sites were blocked using 3% H2O2 (VWR chemicals, 23615.248) and Protein-Block solution (DAKO Agilent technologies, X0909) respectively, for 10 min. Sections were then stained for anti-p16 (ab211542, Abcam, 1:1200), anti-Ki67 (Clone SP6; Lab Vision Corporation #RT-9106-R7, RTU) anti-Phospho-HP1y (Ser83) Antibody (CST #2600, 1:200), anti-CK8 (ab,59400, Abcam), anti-CK5 (ab52635, Abcam). IHC analyses were performed using the Imagescope software.

Bernasocchi T., El Tekle G., Bolis M., Mutti A., Vallerga A., Brandt L.P., Spriano F., Svinkina T., Zoma M., Ceserani V., Rinaldi A., Janouskova H., Bossi D., Cavalli M., Mosole S., Geiger R., Dong Z., Yang C.G., Albino D., Rinaldi A., Schraml P., Linder S., Carbone G.M., Alimonti A., Bertoni F., Moch H., Carr S.A., Zwart W., Kruithof-de Julio M., Rubin M.A., Udeshi N.D, & Theurillat J.P. (2021). Dual functions of SPOP and ERG dictate androgen therapy responses in prostate cancer. Nature Communications, 12, 734.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!