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Antibody mix 2

Manufactured by Novus Biologicals
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Antibody Mix 2 is a combination of multiple antibodies developed for use in laboratory research applications. The core function of this product is to provide a pre-mixed solution of antibodies for convenient and consistent use in various experimental procedures. No further details or interpretation on the intended use of this product can be provided.

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Lab products found in correlation

Antibody mix 1, by Thermo Fisher Scientific (2 mentions) Rat anti cd4, by Thermo Fisher Scientific (2 mentions) Rat anti cd8a, by Thermo Fisher Scientific (2 mentions) Rat anti b220, by BioLegend (2 mentions) Anti rabbit 647, by Cell Signaling Technology (2 mentions) Anti rat 647, by Cell Signaling Technology (2 mentions) Ab15323, by Proteintech (2 mentions)

2 protocols using antibody mix 2

1

Multiparametric Immunostaining of Murine Lung Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sequential immunostaining was performed on 3μm thick Formalin-Fixed Paraffin-Embedded (FFPE) murine lung sections. Briefly, heat mediated antigen retrieval was performed in citrate pH 6.0 (antibody Mix 1, ThermoFisher), or in EDTA pH 9.0 (antibody Mix 2, Novus) buffer. Antibody elution was performed between each staining cycle59 . Antibodies used in Mix 1 were; Rat anti-CD4 (1:200, ThermoFisher #14–9766-82), Rat anti-CD8a (1:200, ThermoFisher #14–0808-82) and Rat anti-B220 (1:500, Biolegend #103202). Antibodies used in Mix 2 were; Rabbit anti-IBA1 (1:1000, VWR #100369–764), Rabbit anti-iNOS (1:100, Abcam #ab15323) and Rabbit anti-CD206 (1:500, ProteinTech #18704–1-AP). Secondary antibodies used were: anti-Rabbit 555 (1:500, Cell Signaling #4413S), anti-Rabbit 647 (1:500, Cell Signaling #4414S) and anti-Rat 647 (1:500, Cell Signaling #4418S).
Acquired images were processed using FIJI and the FIJI plugin HyperStackReg V5.660 (and Ved Sharma. 2018, December 13). ImageJ plugin HyperStackReg V5.6 (Zenodo. http://doi.org/10.5281/zenodo.2252521). Autofluorescence acquired in non-relevant channels was substracted as appropriate. IBA1 and iNOS staining quantitation was performed using Ilastik (v1.3.3post2)61 and CellProfiler (v3.1.9)62 .
Conlon T.M., John-Schuster G., Heide D., Pfister D., Lehmann M., Hu Y., Ertüz Z., Lopez M.A., Ansari M., Strunz M., Mayr C., Ciminieri C., Costa R., Kohlhepp M.S., Guillot A., Günes G., Jeridi A., Funk M.C., Beroshvili G., Prokosch S., Hetzer J., Verleden S.E., Alsafadi H., Lindner M., Burgstaller G., Becker L., Irmler M., Dudek M., Janzen J., Goffin E., Gosens R., Knolle P., Pirotte B., Stoeger T., Beckers J., Wagner D., Singh I., Theis F.J., de Angelis M.H., O’Connor T.O., Tacke F., Boutros M., Dejardin E., Eickelberg O., Schiller H.B., Königshoff M., Heikenwalder M, & Yildirim A.Ö. (2020). Inhibition of LTβR-signalling activates Wnt-induced regeneration in lung. Nature, 588(7836), 151-156.
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2

Multiparametric Immunostaining of Murine Lung Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sequential immunostaining was performed on 3μm thick Formalin-Fixed Paraffin-Embedded (FFPE) murine lung sections. Briefly, heat mediated antigen retrieval was performed in citrate pH 6.0 (antibody Mix 1, ThermoFisher), or in EDTA pH 9.0 (antibody Mix 2, Novus) buffer. Antibody elution was performed between each staining cycle59 . Antibodies used in Mix 1 were; Rat anti-CD4 (1:200, ThermoFisher #14–9766-82), Rat anti-CD8a (1:200, ThermoFisher #14–0808-82) and Rat anti-B220 (1:500, Biolegend #103202). Antibodies used in Mix 2 were; Rabbit anti-IBA1 (1:1000, VWR #100369–764), Rabbit anti-iNOS (1:100, Abcam #ab15323) and Rabbit anti-CD206 (1:500, ProteinTech #18704–1-AP). Secondary antibodies used were: anti-Rabbit 555 (1:500, Cell Signaling #4413S), anti-Rabbit 647 (1:500, Cell Signaling #4414S) and anti-Rat 647 (1:500, Cell Signaling #4418S).
Acquired images were processed using FIJI and the FIJI plugin HyperStackReg V5.660 (and Ved Sharma. 2018, December 13). ImageJ plugin HyperStackReg V5.6 (Zenodo. http://doi.org/10.5281/zenodo.2252521). Autofluorescence acquired in non-relevant channels was substracted as appropriate. IBA1 and iNOS staining quantitation was performed using Ilastik (v1.3.3post2)61 and CellProfiler (v3.1.9)62 .
Conlon T.M., John-Schuster G., Heide D., Pfister D., Lehmann M., Hu Y., Ertüz Z., Lopez M.A., Ansari M., Strunz M., Mayr C., Ciminieri C., Costa R., Kohlhepp M.S., Guillot A., Günes G., Jeridi A., Funk M.C., Beroshvili G., Prokosch S., Hetzer J., Verleden S.E., Alsafadi H., Lindner M., Burgstaller G., Becker L., Irmler M., Dudek M., Janzen J., Goffin E., Gosens R., Knolle P., Pirotte B., Stoeger T., Beckers J., Wagner D., Singh I., Theis F.J., de Angelis M.H., O’Connor T.O., Tacke F., Boutros M., Dejardin E., Eickelberg O., Schiller H.B., Königshoff M., Heikenwalder M, & Yildirim A.Ö. (2020). Inhibition of LTβR-signalling activates Wnt-induced regeneration in lung. Nature, 588(7836), 151-156.
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