Application
IHC - Wholemount
Sample
Zebrafish Embryo (whole embryo - 24hpf)
Specification
whole embryo - 24hpf
Other product details
Dilution
1/200
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: maleic acid blocking solution
Secondary antibody
Name
Non-Abcam antibody was used: AlexaFluor594 Donkey anti-rabbit IgG
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Dilution
1/1000
Additional data
Additional Notes
Whole-mount IHC on 24hpf zebrafish embryos fixed in 4%PFA.
Maleic acid blocking solution is: 150mM maleic acid, 100mM NaCl, 2% blocking reagent (roche), pH7.5, 0.1% Tween-20.
Image shows data for Wnt3a by wholemount IHC of a representative 24hpf zebrafish embryo. When compared to the expression pattern of wnt3a at 24hpf detected by in situ hybridization (as published in Nyholm et al., 2007) this antibody gives high background, but some staining is observed in expected tissue.
Nyholm MK, Wu SF, Dorsky RI and Y Grinblat (2007) The zebrafish zic2a-zic5 gene pair acts downstream of canonical Wnt signaling to control cell proliferation in the developing tectum. Development. 134: 735-746.
Maleic acid blocking solution is: 150mM maleic acid, 100mM NaCl, 2% blocking reagent (roche), pH7.5, 0.1% Tween-20.
Image shows data for Wnt3a by wholemount IHC of a representative 24hpf zebrafish embryo. When compared to the expression pattern of wnt3a at 24hpf detected by in situ hybridization (as published in Nyholm et al., 2007) this antibody gives high background, but some staining is observed in expected tissue.
Nyholm MK, Wu SF, Dorsky RI and Y Grinblat (2007) The zebrafish zic2a-zic5 gene pair acts downstream of canonical Wnt signaling to control cell proliferation in the developing tectum. Development. 134: 735-746.
Abcam response
Thank you for submitting this abreview. Ab28472 has previously been rated 4 star for IHC-wholemount (please see other abreview). However, your data shows that the staining seen does not reflect that of the in situ pattern as publish by Nyholm et al., 2007, under these conditions. Abcam would welcome feedback from other customers using this antibody.
Ms. Emerald Butko
Verified customer
Submitted Mar 09 2012