The following antibodies were purchased: for Western blotting, anti-β-catenin (1:5000; Abcam, cat. no. ab32572), anti-β-actin (1:5000; Bioss, cat. no. bs-0061R), anti-CDK5RAP2 (1:2000; Abcam, cat. no. ab70213), anti-survivin (1:2000; Abclonol, cat. no. A1551), anti-ALDH1 (1:1000; Cell Signaling Technology, cat. no. 54135), anti-Notch1 (1:1000, Abcam, cat. no. ab52627), anti-EZH2 (1:1000; Cell Signaling Technology, cat. no. 5246), anti-CCND1 (1:1000, Cell Signaling Technology, cat. no. 55506), and horseradish peroxidase-conjugated secondary antibodies (1:1000; Cell Signaling Technology, Inc.); for IHC analyses, anti-CDK5RAP2 (1:400; Abcam, cat. no. ab235893), anti-ALDH1 (1:50; Abcam, cat. no. ab52492), anti-SOX2 (1:100; Abcam, cat. no. Ab92494), anti-CD44 (1:4000; Abcam, cat. no. ab189524), anti-CD133 (1:1000; Abcam, cat. no. ab222782), anti-Notch1 (1:150; Abcam, cat. no. ab52627), anti-EZH2 (1:200; Cell Signaling Technology, cat. no. 5246), and anti-CCND1 (1:200; ABclonal, cat. no. A19038); and for immunofluorescence analyses, anti-α-tubulin (1:500, YL1/2; Santa Cruz Biotechnology, cat. no. sc-53029), anti-γ-tubulin (1:1000, GTU88; Sigma-Aldrich, cat. no. T5326), and Alexa Fluor-conjugated secondary antibodies (1:500; Thermo Fisher Scientific).
Anti aldh1
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-ALDH1 is a laboratory reagent used to detect the presence and expression levels of the ALDH1 (Aldehyde Dehydrogenase 1) protein. ALDH1 is an enzyme involved in the oxidation of aldehydes to carboxylic acids. The Anti-ALDH1 product can be used in various analytical techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of ALDH1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti sox2, by Cell Signaling Technology (4 mentions)
Anti cd44, by Abcam (3 mentions)
Anti oct4, by Cell Signaling Technology (3 mentions)
Anti sox2, by Abcam (2 mentions)
Anti bax, by Cell Signaling Technology (2 mentions)
Anti cd44, by Cell Signaling Technology (2 mentions)
Anti cd133, by Abcam (1 mentions)
Anti notch1, by Abcam (1 mentions)
Anti ccnd1, by Cell Signaling Technology (1 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti α tubulin, by Santa Cruz Biotechnology (1 mentions)
Anti γ tubulin, by Merck Group (1 mentions)
Anti cdk5rap2, by Abcam (1 mentions)
Anti aldh1, by Abcam (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Anti ezh2, by Cell Signaling Technology (1 mentions)
Anti β actin, by Bioss Antibodies (1 mentions)
Anti β catenin, by Abcam (1 mentions)
Anti survivin, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
8 protocols using anti aldh1
1
Comprehensive Antibody Panel for Cellular Analyses
2
Protein Expression Analysis in Cancer Cells
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Following the protocols of the manufacturer, RIPA lysis buffer (Beyotime Institute of Biotechnology, Shanghai, China) was used for cellular protein extraction. The BCA Protein Assay Kit (Beyotime Institute of Biotechnology, Shanghai, China) was selected for the detection of protein concentrations. The primary antibodies included rabbit anti-Ki67 antibody (1:25, ab833, Abcam), anti-proliferating cell nuclear antigen (PCNA) (1:1,000, #13110, Cell Signaling), anti-Survivin (1:1,000, #2808, Cell Signaling), anti-Bax (1:1,000, #5023, Cell Signaling), anti-Bcl-2 antibody(1:500, ab196495, Abcam), anti-CD44 (1:100, #37259, Cell Signaling), anti-SOX2 (1:1,000, #14962, Cell Signaling), anti-OCT4 (1:1,000, #2890, Cell Signaling), anti-ALDH1 (1:1,000, #54135, Cell Signaling), β-catenin (1:1,000, #8480, Cell Signaling), c-Myc (1:1,000, #18583, Cell Signaling), and cyclin D1 (1:1,000, #2890, Cell Signaling). Goat anti-rabbit immunoglobin G (IgG) horseradish peroxidase (HRP)-conjugated secondary antibodies were incubated for 1 h at 37 °C. An automatic digital gel image analysis system, Bio-Rad CFX-96 (Bio-Rad, Hercules, CA, USA) was used to determine and analyze band density.
3
Western Blot Analysis of A549 Cells
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A549 cells or tissues were lysed and collected, then balanced with PBS according to Bradford assay. About 40 μg of proteins were used in SDS-PAGE for each sample. Primary and secondary antibodies were incubated according to the manufacturers’ protocols. The proteins of interest were visualized by enhanced chemiluminescence reagents with ChemiDoc XRS. Anti-CD44 (ab157107), anti-Ki67 (ab15580), and anti-Caspase-3 (ab32351) were from Abcam (Cambridge, UK). And anti-ZEB1 (#3396), anti-SOX-2 (#2748), anti-ALDH-1 (#12035), anti-OCT4 (#4286), anti-Bax (#2772), anti-Bcl-2 (#4223), anti-GAPDH (#51332) and all secondary antibodies were from Cell Signaling Technology (Danvers, MA, USA).
4
Western Blotting of Cell Markers
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Procedures for western blotting have been previously described (Li et al., 2018 (link)). Primary antibodies used for western blotting are as follows: anti-GAPDH (Abcam, Cat#: ab9484, 1:5,000), anti-CD44 (Abcam, Cat#: ab157107, 1:5,000), anti-PER3 (Abcam, Cat#: ab177482, 1:5,000), anti-ALDH1 (Cell Signaling Technology, Cat#:5741, 1:5,000), anti-total β-catenin (Cell Signaling Technology, Cat#:2951, 1:5,000), anti-phosphorylated β-catenin (Cell Signaling Technology, Cat#: 176, 1:5,000).
5
Protein Expression in Colorectal Cancer
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Protein samples extracted from colorectal cancer cells after indicated treatments were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes. After blocking in 5% nonfat milk, the membranes were incubated with anti-CXCR4 (1:1,000; Abcam, Cambridge, MA, USA), anti-SOX2 (1:1,000), anti-FOXM1 (1:1,000), anti-CD133 (1:1,000), anti-ALDH1 (1:1,000), anti-OCT4 (1:1,000), anti-CD44 (1:1,000; Cell Signaling Technology, Danvers, MA, USA), and anti-β-actin (1:5,000; Proteintech, Rosemont, IL, USA) antibodies overnight at 4°C. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies. Protein signals were detected by enhanced chemiluminescence.
6
Protein Expression Analysis Protocol
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Cells were lysed using radioimmunoprecipitation assay (RIPA) buffer (Solarbio, Cat# R0010) and added with a cocktail of protease inhibitors and phosphatase inhibitors (Huaxingbio, Chaohu, China; Cat# HX1864). Total proteins (30 μg) were separated using SDS polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto polyvinylidene fluoride membranes. The membranes were blocked with 5% skim milk for 1 h and then incubated with primary antibodies. The primary antibodies used in this study were: anti-MYC (Cell Signaling Technology; Cat# 18583; 1:1000), anti-B lymphoma Mo-MLV insertion region 1 homolog (BMI1) (Cell Signaling Technology; Cat# 6964; 1:1000), anti-SRY-box transcription factor 2 (SOX2) (Cell Signaling Technology; Cat# 14962; 1:1000), anti‑ALDH1 (Cell Signaling Technology; Cat# 54135; 1:1000), anti-phospho-Histone H2A.X (Cell Signaling Technology; Cat# 9718; 1:1000), anti-p-IRF3 (Cell Signaling Technology; Cat# 37829; 1:1000), anti-GAPDH (ZSGB-BIO; Cat# TA-08; 1:1000), and anti-Histone H3 (ABclonal, Wuhan, China; Cat# A2348; 1:1000). The membranes were then stained with the appropriate secondary antibodies. The signals were detected using a Clarity Western ECL kit (Thermo Fisher Scientific; Cat# 34577).
7
Antibodies for Cancer Stem Cell Analysis
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The following antibodies were used in this study: rabbit monoclonal anti‐TRAF6, anti‐CD44, anti‐SOX2 and anti‐C4.4A (Abcam, Cambridge, UK); mouse monoclonal anti‐CD44 (Abcam); rabbit monoclonal anti‐CD44, anti‐KLF4 and anti‐SOX2 (Epitomics, Burlingame, CA, USA); rabbit polyclonal anti‐ALDH1 (GeneTex Inc., Irvine, CA, USA); rabbit monoclonal anti‐Vimentin, anti‐N‐cadherin, anti‐E‐cadherin, anti‐Slug, anti‐NF‐κB p65, anti‐Phospho‐NF‐κB p65 and anti‐ALDH1 (Cell Signaling Technology, Boston, MA, USA); rabbit polyclonal anti‐AGR2 (Cell Signaling Technology).
8
Stem Cell Marker Expression Analysis
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Primer sequences (bp)
Plus SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific, Waltham, MA, USA) for 60 sec. Protein levels were normalized with respect to the band density of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control. The primary antibodies used were: anti-ABCG2 (1:1,000), anti-Bmi-1 (1:1,000), anti-c-Myc (1:1,000), anti-Nanog (1:1,000), anti-Oct3/4 (1:1,000), anti-Sox2 (1:1,000), anti-STAT3 (1:1,000), and anti-ALDH1 (1:1,000) (all from Cell Signaling Technology), and anti-GAPDH (1:5,000) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, uSA).
Statistical analysis. The statistical software SPSS 20.0 (SPSS Inc., Chicago, IL, uSA) was used for data processing and analyzing. Data are presented as the means ± SD, and a comparison was carried between experimental groups of qPCR analysis using one-way ANOVA. The results of the chemosensitivity assay were calculated and compared using one-way ANOVA. P<0.05 was considered statistically significant.
Plus SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific, Waltham, MA, USA) for 60 sec. Protein levels were normalized with respect to the band density of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control. The primary antibodies used were: anti-ABCG2 (1:1,000), anti-Bmi-1 (1:1,000), anti-c-Myc (1:1,000), anti-Nanog (1:1,000), anti-Oct3/4 (1:1,000), anti-Sox2 (1:1,000), anti-STAT3 (1:1,000), and anti-ALDH1 (1:1,000) (all from Cell Signaling Technology), and anti-GAPDH (1:5,000) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, uSA).
Statistical analysis. The statistical software SPSS 20.0 (SPSS Inc., Chicago, IL, uSA) was used for data processing and analyzing. Data are presented as the means ± SD, and a comparison was carried between experimental groups of qPCR analysis using one-way ANOVA. The results of the chemosensitivity assay were calculated and compared using one-way ANOVA. P<0.05 was considered statistically significant.
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