The immunohistochemistry technique of the present study followed the protocol of the International Research Center of the A.C.Camargo Cancer Center. Briefly, the 3 μm tissue sections were dewaxed and dehydrated. For the ALDH1 antibody, antigen retrieval was performed with citrate solution (10 mM, pH = 6.0) and for Notch1 antibody, the EDTA solution (10 m, pH = 8.0) was used, both at 110°C, in a pressurized chamber for 15 minutes. The endogenous peroxidase and nonspecific protein activity were blocked with 3% aqueous hydrogen peroxide and Protein Block Serum-Free (Dako, Carpinteria, CA, USA), respectively, followed by incubation with the primary antibodies in a humid chamber (4°C-18 hours): anti-ALDH1 antibody (clone 44, BD Biosciences, San Jose, CA, USA, 1 : 500) and anti-Notch1 antibody (clone D1E11, Cell Signaling Technology, Beverly, MA, USA, 1 : 200). After revelation with adequate chromogenic substrate reaction of 3,3′-diaminobenzidine tetrahydrochloride (DAB, Dako, Carpinteria, CA, USA), the sections were stained with Harris Hematoxylin.
Anti aldh1 antibody clone 44
Manufactured by BD
Sourced in United States
The Anti-ALDH1 antibody (clone 44) is a laboratory reagent used in research applications. It is designed to detect and bind to the ALDH1 (Aldehyde Dehydrogenase 1) protein, which is a member of the aldehyde dehydrogenase enzyme family. The antibody can be used in various immunoassay techniques to study the expression and localization of ALDH1 in biological samples.
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2 protocols using anti aldh1 antibody clone 44
1
Immunohistochemical Staining for ALDH1 and Notch1
2
Immunohistochemical Evaluation of ALDH1
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ALDH1 expression was assessed using a mouse monoclonal anti-ALDH1 antibody (clone 44, 1:200 dilution; BD Biosciences, San Jose, CA) by immunohistochemistry (IHC). Briefly, tissue samples (5 μm) were fixed in 4% formaldehyde and embedded in paraffin. The sections were then treated with Target Retrieval (0.01 mol/L; pH 6.0) under high pressure using a microwave for 15 min. Endogenous peroxidase activity was blocked by Dako Dual Endogenous Enzyme Block (Dako, Carpinteria, CA) for 10 min. The sections were subsequently incubated with an anti-ALDH1A1 antibody for 30 min at room temperature, followed with a horseradish peroxidase-labeled rabbit anti-mouse secondary antibody. The sections were washed with phosphate-buffered saline (PBS) and incubated with Envision+ Rabbit Polymer (cat. no. K4003; Dako) for 30 min. The staining was then visualized using 200 μL DAB plus (Dako) chromogen for 10 min and counterstained with Mayer's hematoxylin-eosin (magnification ×400). Negative controls were performed by substituting the primary antibody with either PBS or isotype-specific IgG controls.
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