Cells were attached to chambered slides overnight. Cells were fixed in 10% formalin for 30 minutes, rinsed for 3 times with PBS, blocked and permeabilized with blocking buffer (PBS with 5% BSA, 1% TWEEN®20 and 0.1% Trito-X100, Sigma-Aldrich, St. Louis, MO) for 1 hour at room temperature and incubated for 1 hour with the primary antibody diluted in blocking buffer. Primary antibodies were used as follows: MITF anti mouse (EMD Millipore, Billerica, MA) 1:100; TFE3 anti rabbit (Sigma-Aldrich) 1:200; TFEB anti rabbit (Bethyl Laboratories, Montgomery, TX) 1:100. Following three 5-minute washes in PBST (PBS with 0.2% TWEEN®20) cells were incubated for 1 hour at room temperature with secondary antibodies Alexa Fluor 488 anti-mouse IgG (1:400) and Alexa Fluor 594 anti-rabbit IgG (1:400) (Thermo Fisher Scientific, Waltham, MA). Cell nuclei were counterstained with DAPI (0.1 μg/ml) for 15 minutes in PBST and cells were washed 3 times in PBST before mounting the slides with antifade mounting medium (Vector Laboratories, Burlingame, CA). Images were acquired with a fluorescence microscope (BZ-X700 Keyence, Osaka, JN).
Tfe3 anti rabbit
Manufactured by Merck Group
TFE3 anti rabbit is a laboratory reagent used in immunological techniques, such as western blotting and immunohistochemistry. It is a polyclonal antibody that specifically binds to the TFE3 protein, which is a transcription factor involved in various cellular processes. The primary function of this product is to enable the detection and analysis of TFE3 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Human osteoactivin gpnmb anti goat, by R&D Systems (3 mentions)
Mouse anti mitf, by Merck Group (2 mentions)
Rabbit anti tfeb, by Fortis Life Sciences (2 mentions)
Axio scan z1 slide scanner, by Zeiss (2 mentions)
Antifade mounting medium, by Vector Laboratories (1 mentions)
Alexa fluor 488 anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Bz x700, by Keyence (1 mentions)
Melan a, by Abcam (1 mentions)
Cytokeratin 7, by Abcam (1 mentions)
Cathepsin k, by Abcam (1 mentions)
Pan cytokeratin, by Novus Biologicals (1 mentions)
Hematoxylin, by Vector Laboratories (1 mentions)
4 protocols using tfe3 anti rabbit
1
Immunofluorescence Staining of MITF, TFE3, and TFEB
2
Immunohistochemical Analysis of Kidney Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hematoxylin and eosin staining was performed by standard methods. Histology was reviewed by a pathologist experienced in evaluating kidney cancer. Immunohistochemistry for TFE3 and GPNMB was performed as previously described [28 ]. Primary antibodies were as follows: Human Osteoactivin/GPNMB anti goat (R&D Systems; 1:800); TFE3 anti rabbit (Sigma Aldrich; 1:800). Spectral karyotyping and TFE3 was performed as previously described [29 (link)] and outlined in Supplementary Methods.
3
Immunohistochemical Profiling of Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Five-micron thick formalin-fixed paraffin-embedded sections were cut from all available tumor blocks. IHC staining and H&E staining were performed by either VitroVivo Biotech (Rockville, MD) or the Molecular Histopathology Laboratory at the Frederick National Laboratory for Cancer Research (Frederick, MD) using standardized methodologies. IHC staining was performed using either Human Osteoactivin/GPNMB anti-goat (R&D Systems, Minneapolis, MN), TFE3 anti-rabbit (Sigma Aldrich, Burlington, MA), Phospho-S6 Ribosomal Protein (Ser235/236) anti-rabbit (Cell Signaling Technology, Danvers, MA), or Phospho-4-E-BP1 (Thr37/46) anti-rabbit (Cell Signaling Technology, Danvers, MA). For selected tumors, an immunohistology panel was performed evaluating HMB45 (Agilent Dako, Carpinteria, CA), Melan-A (Abcam, Waltham, MA), Pan-cytokeratin (Novus Biologicals, Centennial, CO), Cytokeratin 7 (Abcam, Waltham, MA), Cathepsin K (Abcam, Waltham, MA), and Carbonic anhydrase IX (Cell Signaling Technology, Danvers, MA). Images were captured using an AxioScan.Z1 Slide Scanner (Zeiss, Oberkochen, DE).
4
Immunohistochemistry of Melanoma Markers
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!