CHD7 antibody | knockout validation | Cell Signaling 6505

This is a knockout-validated antibody summary, based on the publications "Chd7 is indispensable for mammalian brain development through activation of a neuronal differentiation programme", as cited below [1] and "CHARGE syndrome protein CHD7 regulates epigenomic activation of enhancers in granule cell precursors and gyrification of the cerebellum" for (figure s1b,s1c) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

CHD7 antibody | knockout validation | Cell Signaling 6505 figure 1
Figure 1. Western blot showing the depletion of CHD7 in CHD7-KO HEK293T cells. RPA116 was used as a loading control. From [1].
Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: CHD7

Catalog number: 6505

Summary: Rabbit monoclonal IgG against a recombinant protein specific to the amino terminus of human CHD7 protein. Reacts with human and mouse by western blot and immunoprecipitation.

CHD7 antibody | knockout validation | Cell Signaling 6505 figure 2
Figure 2. (a) Immunostaining of Chd7 in P7 cerebella of Chd7 WT (Chd7f/f) and homozygous mutant (Atoh1-Cre::Chd7f/f). DNA was stained with DAPI. Scale bars, 50 um. EGL: external granule layer. IGL: internal granule layer. (b) Immunostaining of Chd7 (in green) in isolated WT (Chd7f/f) and mutant (Atoh1-Cre::Chd7f/f) cerebellar granule progenitors (GNPs) cultured without Shh for 48 hours. Note that most of mutant cells lost Chd7 staining. Scale bars, 50 um. From [1].
Validation Method

Western blot | Immunohistochemistry | Immunocytochemistry

Sample

WT and CHD7-KO mice cerebellum and cells (cerebellar granule progenitors and HEK293T). IHC: animals were killed and fixed with 4% paraformaldehyde in PBS (PFA/PBS) overnight, followed by sectioning with cryostat (Leica) and microtome (Leica). IC: cells were fixed with 4% PFA/PBS for 10 min at room temperature.

Blocking agent

IC/IHC: 10% normal donkey serum.

Primary incubation

IC/IHC: 1:400 dilution overnight at 4 °C .

Detection

IC/IHC: Fluorescent images were captured using a confocal laser-scanning microscopy (LSM780 and 800, Zeiss; and SP5, Leica)

References
  1. Feng W, Kawauchi D, Körkel Qu H, Deng H, Serger E, Sieber L, et al. Chd7 is indispensable for mammalian brain development through activation of a neuronal differentiation programme. Nat Commun. 2017;8:14758 pubmed publisher
  2. Reddy N, Majidi S, Kong L, Nemera M, Ferguson C, Moore M, et al. CHARGE syndrome protein CHD7 regulates epigenomic activation of enhancers in granule cell precursors and gyrification of the cerebellum. Nat Commun. 2021;12:5702 pubmed publisher