Abcam Monoclonal Egr-1 ab133695 is an effective primary antibody replacement for Santa Cruz sc-189 polyclonal Egr-1 in songbirds

Erin N Scully; Juliana Montoya Sanchez; Christopher B Sturdy

doi:10.1016/j.heliyon.2019.e02938

Abcam Monoclonal Egr-1 ab133695 is an effective primary antibody replacement for Santa Cruz sc-189 polyclonal Egr-1 in songbirds

Heliyon. 2019 Nov 28;5(11):e02938. doi: 10.1016/j.heliyon.2019.e02938. eCollection 2019 Nov.

Authors

Erin N Scully¹, Juliana Montoya Sanchez¹, Christopher B Sturdy^{1

2}

Affiliations

¹ Department of Psychology, University of Alberta, Canada.
² Neuroscience and Mental Health Institute, University of Alberta, Canada.

Abstract

Background: The immediate early gene ZENK (acronym zif268, Egr-1, NGFI-A, krox24) has been used extensively in songbird research (Mello et al., 1992; Jarvis and Nottebohm, 1997), as well as other research areas. ZENK has been used in assessing learning and memory, measuring neural activation, and identifying the cellular and molecular substrates involved in the first stages of memory formation (Watson and Clements, 1980). Previous songbird research has found that neurons located within the areas involved in auditory perception, namely the caudomedial nidopallium and caudomedial mesopallium, exhibit high levels of ZENK protein expression in response to conspecific songs and calls (Mello and Ribeiro, 1998; Avey et al., 2011).

New method: In large part due to its neuronal-specific labeling of ZENK protein, Santa Cruz Egr-1 sc-189 has been widely accepted as the standard primary antibody in songbird research. However, Santa Cruz Biotechnology Egr-1 no longer specifically labels and has also discontinued production of Egr-1 sc-189. Thus, the current study is focused on analyzing the effectiveness of alternative primary antibodies: Abcam polyclonal c-Fos, Abcam monoclonal ab133695 Egr-1, and Proteintech polyclonal Egr-1.

Results: Abcam monoclonal Egr-1 was successful in specifically labeling ZENK positive cells in the songbird auditory nuclei. Abcam polyclonal c-Fos and Proteintech polyclonal Egr-1 were found to have non-specific labeling.

Comparison with existing methods: Abcam monoclonal Egr-1 ab133695 was found to produce differential and specific labeling in the targeted auditory nuclei similar to previous studies successfully using Santa Cruz polyclonal Egr-1 (i.e. Mello and Ribeiro, 1998).

Conclusions: Abcam monoclonal Egr-1 effectively labels ZENK in the songbird auditory nuclei, making it a suitable primary antibody replacement for Santa Cruz polyclonal Egr-1.

Keywords: Behavioral neuroscience; Cellular neuroscience; Egr-1; Immunohistochemistry; Nervous system; Neuroanatomy; Neuroscience; Songbird; ZENK.