Mucin MUC5AC antibody | knockout validation | Millipore MAB2011

This is a knockout-validated antibody summary, based on the publication "Molecular Implications of MUC5AC-CD44 Axis in Colorectal Cancer Progression and Chemoresistance", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mucin MUC5AC antibody | knockout validation | Millipore MAB2011 figure 1
Figure 1. Immunoblot showing deletion of MUC5AC in clones of CRC HCT-8 cell line. From [1].
Antibody information

Mouse monoclonal IgG1

Company: Millipore

Antibody: Mucin MUC5AC

Catalog number: MAB2011

Summary: Mouse monoclonal IgG1 against a synthetic peptide of the MUC5AC tandem repeat. Reacts with human by western blot, immunoprecipitation, immunocytochemistry, immunohistochemistry (paraffin) and ELISA.

Mucin MUC5AC antibody | knockout validation | Millipore MAB2011 figure 2
Figure 2. Immunoblot showing deletion of MUC5AC in clones of CRC LS174T cell lines. From [1].
Validation Method

Western blot

Sample

WT and MUC5AC-KO HCT-8 and LS174T cell lines. Total protein lysates were prepared in radio immunoprecipitation assay (RIPA) lysis buffer containing 50 mM Tris-HCl, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate along with protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany).

Blocking agent

5% non-fat dry milk having phosphate-buffered saline with 0.1% Tween 20 (PBST) for 1 h.

Primary incubation

At 4 °C overnight.

Secondary incubation

For 1 h at room temperature.

Detection

Chemiluminescence reagent (GE Healthcare Bio-Sciences, PA, USA).

Clone note

The same clone (CLH2) is sold as Santa Cruz Biotechnology sc-33667; LifeSpan Biosciences LS-B2434, LS-C152909, LS-C312111, LS-C348684; Abnova MAB13373, MAB13374; EMD Millipore MAB2011.

References
  1. Pothuraju R, Rachagani S, Krishn S, Chaudhary S, Nimmakayala R, Siddiqui J, et al. Molecular implications of MUC5AC-CD44 axis in colorectal cancer progression and chemoresistance. Mol Cancer. 2020;19:37 pubmed publisher