Ab75869

Manufactured by Abcam

Sourced in United Kingdom

Ab75869 is a laboratory equipment product offered by Abcam. It serves a core function as a measurement and analysis tool, however, further details on its specific intended use or capabilities are not available at this time.

Automatically generated - may contain errors

Lab products found in correlation

Ab109725, by Abcam (2 mentions) Ab5694, by Abcam (2 mentions) Ab86734, by Abcam (1 mentions) A11037, by Thermo Fisher Scientific (1 mentions) A11032, by Thermo Fisher Scientific (1 mentions) Immpress peroxidase detection kit, by Vector Laboratories (1 mentions) A11008, by Thermo Fisher Scientific (1 mentions) A11029, by Thermo Fisher Scientific (1 mentions) Ab24479, by Abcam (1 mentions) Sc 11418, by Santa Cruz Biotechnology (1 mentions) Nanozoomer, by Hamamatsu Photonics (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Ab15580, by Abcam (1 mentions) Ab24590, by Abcam (1 mentions) Sf solution, by Merck Group (1 mentions) Ab187646, by Abcam (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Dab substrate kit, by Agilent Technologies (1 mentions) Permount mounting medium, by Thermo Fisher Scientific (1 mentions) Ab68459, by Abcam (1 mentions)

6 protocols using ab75869

Immunostaining Protocols for Cell and Tissue Analysis

Cited 2 times

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

Immunocytochemistry (ICC) was essentially as described (19 (link)). Primary antibodies as noted above CK5 (1:100), CK17 (1:200); vimentin (1:200), or CK8/18 (1:2000) for 2 h, secondary antibodies (A11029, A11037, A11008, A11032, Invitrogen, 1:200) for 1 h, and counterstained with DAPI. Cells were imaged using the Olympus BX40 fluorescent microscope and channels merged in Adobe Photoshop 2021. Immunohistochemistry (IHC) was performed as previously described (33 (link)) using CK5 (rabbit, ab75869, Abcam, 1:200), CK17 (rabbit, ab109725, Abcam, 1:100), vimentin (mouse, 5G3F10, Cell Signaling Technologies, 1:100), or pan-CK (mouse ab86734, 1:400, abcam, Waltham, MA) antibodies and either fluorescent secondary antibodies as described and imaged using an Olympus BX40 fluorescent microscope, or ImmPRESS Peroxidase detection kit (Vector Laboratories).

McGinn O., Riley D., Finlay-Schultz J., Paul K.V., Kabos P, & Sartorius C.A. (2022). Cytokeratins 5 and 17 maintain an aggressive epithelial state in basal-like breast cancer. Molecular cancer research : MCR, 20(9), 1443-1455.

+ Open protocol

+ Expand

Immunohistochemical Analysis of IPF Lung Tissue

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

ZytoChem-Plus AP Kit (Fast Red) (Zytomed Systems, Berlin, Germany) was used for immunohistochemical localization of research target-proteins in formalin-fixed, paraffin-embedded lung tissue sections from patients with sporadic IPF (n = 5) and organ donors (n = 5), according to the manufacturer´s instructions and previous published work [31 (link)]. In the following, the primary antibodies used for IHC are listed, including the sources and dilutions: rabbit polyclonal for human alpha-smooth muscle actin [α-SMA] (1:200, Abcam, ab5694), rabbit monoclonal for human cytokeratin-5 [KRT5] (1:200, Abcam, ab75869), rabbit polyclonal for human survivin (1:200, Abcam, ab24479), mouse monoclonal for human phospho-STAT3 [Y705] (1:25, Cell Signaling Technology, #4113S) and rabbit polyclonal for human HDAC4 (1:50, Santa Cruz, sc-11418). As control experiments, the first antibody was omitted on some sections during staining procedures. Immunostained lung sections were scanned with a scanning device (Nano-Zoomer, Hamamatsu), and examined histopathologically using the ´NDP.view2 software´ at 50×, 100×, 200× and 400× original magnification.

Korfei M., Stelmaszek D., MacKenzie B., Skwarna S., Chillappagari S., Bach A.C., Ruppert C., Saito S., Mahavadi P., Klepetko W., Fink L., Seeger W., Lasky J.A., Pullamsetti S.S., Krämer O.H, & Guenther A. (2018). Comparison of the antifibrotic effects of the pan-histone deacetylase-inhibitor panobinostat versus the IPF-drug pirfenidone in fibroblasts from patients with idiopathic pulmonary fibrosis. PLoS ONE, 13(11), e0207915.

+ Open protocol

+ Expand

Extracellular Matrix Characterization Protocol

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

HA (Mw ≈ 180 kDa, MB7259‐1) was obtained from Meilunbio®. SF solution (Mw ≈ 100 kDa, 5154) was obtained from Sigma. Hexafluoroisopropanol (HFIP, 920‐66‐1) was purchased from Aladdin. Dulbecco's modified Eagle's medium (DMEM, 10567022), fetal bovine serum (FBS, 12483020), penicillin/streptomycin (P/S, 10378016), trypsin (0.25%, 15,050,057), phosphate‐buffered saline (PBS, pH 7.4, 10,010,023), Rhodamine‐ and Alexa Fluor‐488 conjugated phalloidin (R415 and A12379), DAPI (62248) were purchased from Gibco. Primary antibodies to cytokeratin‐5 (K5, ab75869), uroplakin‐3 (ab187646), α‐smooth muscle actin (α‐SMA, ab5694), CD31 (ab24590), and Ki67 (ab15580), and secondary antibodies for fluorescence staining were purchased from Abcam.

Niu Y., Galluzzi M., Deng F., Zhao Z., Fu M., Su L., Sun W., Jia W, & Xia H. (2021). A biomimetic hyaluronic acid‐silk fibroin nanofiber scaffold promoting regeneration of transected urothelium. Bioengineering & Translational Medicine, 7(2), e10268.

+ Open protocol

+ Expand

Immunostaining of CK5 and CK17 in Tissue

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

Tissue sections were processed as above and were stained with antibodies specific for CK5 (rabbit, 1:200, ab75869, Abcam, Cambridge, UK) and CK17 (rabbit, 1:100, ab109725, Abcam) for 2 h at RT. Sections were blocked using HRP Blocking Reagent (Abcam) EnVision+/HRP Visualization (Agilent, Santa Clara, CA, USA) and DAB substrate kit (Agilent) were used to visualize staining. Sections were counterstained with hematoxylin and mounted with Permount Mounting Medium (Fisher Scientific, Waltham, MA, USA). Representative photographs were taken under a light microscope at ×20 magnification.

Li Z., McGinn O., Wu Y., Bahreini A., Priedigkeit N.M., Ding K., Onkar S., Lampenfeld C., Sartorius C.A., Miller L., Rosenzweig M., Cohen O., Wagle N., Richer J.K., Muller W.J., Buluwela L., Ali S., Bruno T.C., Vignali D.A., Fang Y., Zhu L., Tseng G.C., Gertz J., Atkinson J.M., Lee A.V, & Oesterreich S. (2022). ESR1 mutant breast cancers show elevated basal cytokeratins and immune activation. Nature Communications, 13, 2011.

+ Open protocol

+ Expand

Immunohistochemical Analysis of IPF Lung Sections

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

Immunohistochemistry (IHC) of the lung sections was performed as previously described in detail²⁰. The primary antibodies used for IHC are listed as follows: proSP-C rabbit polyclonal antibody (1:750, Merck Millipore, AB3786), rabbit monoclonal antibody for human cytokeratin-5 (KRT5) (1:200, Abcam, Cambridge, United Kingdom, ab75869), rabbit monoclonal antibody for human cytokeratin-7 (KRT7) (1:200, Abcam, ab68459), HOPX mouse monoclonal antibody (1:50, Santa Cruz Inc., sc-398703), and mouse monoclonal antibody for human Ki67 (1:50, Dianova, DIA-670-P05). Immunostained lung sections were scanned with a scanning device (Nano-Zoomer, Hamamatsu), and examined histopathologically using the ‘NDP.view2 software’ at 200×, 400× and 800× original magnification. IHC for mentioned antibodies was undertaken in 12 IPF- and 5 control-donor lung samples.

Ota C., Ng-Blichfeldt J.P., Korfei M., Alsafadi H.N., Lehmann M., Skronska-Wasek W., M. De Santis M., Guenther A., Wagner D.E, & Königshoff M. (2018). Dynamic expression of HOPX in alveolar epithelial cells reflects injury and repair during the progression of pulmonary fibrosis. Scientific Reports, 8, 12983.

+ Open protocol

+ Expand

Immunocytochemistry of Cancer Cells

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

ICC experiments were performed by fixing cells to glass cover slips in ice cold 70% acetone/30%methanol for 5 minutes followed by immunostaining with the indicated antibodies. Antibodies used were rabbit mAb to ERα (SP1); mouse mAb to CK5 (XM26); rabbit mAb to CK5 (ab75869, Abcam, Cambridge, MA); rabbit mAb to BrdU (ab152095, abcam, Cambridge, MA); and rabbit pAB to cleaved caspase-3 (G748A, Promega, Madison, WI). Secondary antibodies utilized included anti-mouse Alexa Fluors 594 (red) and 488 (green), as well as anti-rabbit Alexa Fluors 594 (red) and 488 (green) (Thermo Scientific). For BrdU experiments, cells were treated with 0.25 mg/mL BrdU labeling reagent for 1 hour prior to fixation. Cell nuclei were counterstained with DAPI. All experiments were performed in biological triplicate averaging 5 high power field images per slide. An Olympus CKX41 microscope was used for photography and image analysis. Images were taken in black and white using cellSens software (Olympus) and merged in Adobe Photoshop (Adobe).

Corr B.R., Finlay-Schultz J., Rosen R.B., Qamar L., Post M.D., Behbakht K., Spillman M.A, & Sartorius C.A. (2015). Cytokeratin 5 positive cells represent a therapy resistant subpopulation in epithelial ovarian cancer. International journal of gynecological cancer : official journal of the International Gynecological Cancer Society, 25(9), 1565-1573.

+ 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!