CD24 antibody | knockout validation | Santa Cruz sc-19651

This is a knockout-validated antibody summary, based on the publication "CD24 expression does not affect dopamine neuronal survival in a mouse model of Parkinson's disease", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

CD24 antibody | knockout validation | Santa Cruz sc-19651 figure 1
Figure 1. (A) Strong CD24 expression in the olfactory bulb (especially the glomerular layer and rostral migratory stream). (B) In the entire lining of the ventricular zone at the level of the striatum, CD24 is present. (C) CD24 protein is observed the Reticular nucleus of the thalamus (RT). (D) At the level of the thalamus, strong CD24 expression is seen in the amygdala and the lateral posterior thalamic nucleus (mediocaudal part; LPMC). (E) The LPMC expression is maintained into the midbrain, where there is also very strong staining in the zonal layer and weaker expression in the ventral layers of the superior colliculus (SC). (F) The SC expression of CD24 is maintained throughout the midbrain. There is also CD24 staining surrounding the medial geniculate nucleus (MG). (G) Robust expression in the red nucleus. (H) In the hindbrain, there is strong expression in the nucleus of lateral lemniscus (NLL) and the superior olivary complex (SOC). (I-L) CD24 expression on sections of brain from the Cd24-/- mouse, demonstrating a lack of protein labelling at all levels of the CNS. Numbers on the periphery represent the plane of the coronal section relative to bregma. Scale bar in panel D represents 3mm in images A-D and I-L, and 1.5mm for images E-H. Please see Figure S2 in the article [1].
Antibody information

Rat monoclonal IgG2b

Company: Santa Cruz

Antibody: CD24

Catalog number: sc-19651

Summary: Rat monoclonal IgG2b against C57BL/10 mouse splenic T cells and concanavalin A-activated splenocytes. Recommended for detection of mouse CD24 by western blot, immunoprecipitation, immunofluorescence, immunohistochemistry (paraffin) and flow cytometry.

Validation Method

Immunohistochemistry

Sample

Brain from control and Cd24-/- mouse. PFA fixed brains were shifted to 30% sucrose, while the formalin fixed brains were washed in PBS and switched to 70% ethanol for paraffin embedding. Immunohistochemistry was carried out with free-floating sections.

Primary incubation

1:50 dilution in 5% normal horse serum plus 0.05% Triton X-100 in PBS at room temperature overnight.

Secondary incubation

1:200 dilution with an appropriate biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) for one-hour.

Detection

40-minute incubation in avidin-biotin-peroxidase solution (ABC Elite; Vector Laboratories) followed by 3’-diaminobenzidine (DAB; Sigma).

References
  1. Stott S, Hayat S, Carnwath T, Garas S, Sleeman J, Barker R. CD24 expression does not affect dopamine neuronal survival in a mouse model of Parkinson's disease. PLoS ONE. 2017;12:e0171748 pubmed publisher