Anti-TBR1 抗体

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TBR1 antibody (AB31940)

IHC image of TBR1 staining in a section of formalin-fixed paraffin-embedded normal E17 mouse brain performed on a Leica BOND^TM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab31940, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TBR1 antibody (AB31940)

(Image courtesy of Human Protein Atlas)
ab31940 staining TBR1 protein in normal human cerebral cortex. Brown color indicates presence of protein, blue color shows cell nuclei. Paraffin embedded human cerebral cortex tissue was incubated with ab31940 at a 1/25 dilution for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

Image courtesy of Human Protein Atlas

• IHC-P

AbReview32152****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TBR1 antibody (AB31940)

IHC-P image of TBR1 staining on mouse brain sections using ab31940 at a 1 : 200 dilution.

The sections were deparaffinized and subjected to heat mediated antigen retrieval. The sections were blocked using 7.5% goat serum for 2 hours at room temperature. ab31940 was diluted 1 : 200 using blocking buffer and incubated with the sections for 16 hours at 4°C. The secondary antibody used was Goat polyclonal to anti-rabbit conjugated to Alexa Fluor^® 594 (1 : 400).

DAPI was used to counterstain nuclei.

This image is courtesy of an anonymous abreview.

• WB

Lab

Western blot - Anti-TBR1 antibody (AB31940)

Gel type : MOPS

Blocking buffer : 3% milk block

All lanes:

Western blot - Anti-TBR1 antibody (ab31940) at 1 µg/mL

Lane 1:

Mouse hippocampus tissue lysate at 20 µg

Lane 2:

Rat hippocampus tissue lysate at 20 µg

Lane 3:

Human hippocampus tissue lysate at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

Predicted band size: 74 kDa

Observed band size: 74 kDa

false

Exposure time: 4min

• WB

Lab

Western blot - Anti-TBR1 antibody (AB31940)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab31940 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-TBR1 antibody (ab31940) at 1 µg/mL

Lane 1:

Mouse hippocampus whole cell lysate at 20 µg

Lane 2:

Rat hippocampus whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Predicted band size: 74 kDa

Observed band size: 50 kDa,74 kDa

true

Exposure time: 4min

• IHC

CiteAb

Immunohistochemistry - Anti-TBR1 antibody (AB31940)

Immunohistochemistry-immunofluorescence using Anti-TBR1 antibody, ab31940. Publication image from Hattori, Y. et al., 2020, Nat Commun, 32242005. Legend direct from paper.

In vivo administration of anti-IL-6 and/or IFNAR1 antibodies significantly attenuated the disturbed expression of neuronal subtype-associated transcription factors in postmigratory neurons.a The experimental design for the administration of neutralizing antibodies, which is combined with in vivo CXCL12 overexpression in postmigratory neurons. b Representative immunostaining (shown in pseudo color) for GFP (CX3CR1; red), Tbr1/Ctip2/Satb2/Cux1 (green), and DAPI (blue) in control and CXCL12-overexpressing brains that were mock-treated or administered neutralizing antibodies for IL-6 or IFNAR1. Scale bar, 50 µm. c The density of cells expressing Ctip2, Satb2, Cux1, or Tbr1 per total (DAPI+) cells in 20 µm bins in the CP, which was divided into eight zones (two-sided Steel–Dwass test; n = 8 sections from four mice; **P < 0.01, *P < 0.05). Data are presented as the mean values ± S.D. The exact P values are summarized in Supplementary Table 1. Source data are provided as a Source Data File.

• IHC

CiteAb

Immunohistochemistry - Anti-TBR1 antibody (AB31940)

Immunohistochemistry-immunofluorescence using Anti-TBR1 antibody, ab31940. Publication image from Hattori, Y. et al., 2020, Nat Commun, 32242005. Legend direct from paper.

In vitro-prepared CP-like neurons changed subtype marker expression upon exposure to microglia.a Immunostaining for d4Venus (Gadd45g; green), Tbr2 (red), MAP2 (cyan), and DAPI (blue) in E14 Gadd45g-d4Venus Tg mouse cerebral walls. The broken lines indicate apical and basal surfaces. Scale bar, 50 µm. b The protocol for the in vitro preparation of CP-like neurons, which were obtained from Gadd45g-d4Venus+ cells isolated from the E14 Gadd45g-d4Venus cortex by FACS, after culture for 2 days. c A time-lapse series showing the reduction in Gadd45g-dVenus expression during culture. Scale bar, 20 µm. d CX3CR1-GFP+ microglia harvested from E15 CX3CR1-GFP pallial walls by FACS were added to the CP-like neuronal cultures. e Representative immunostaining (shown in pseudo color) for βIII-tubulin (cyan), Tbr1/Satb2/Cux1 (red), and Ctip2 (green). The arrowhead indicates microglia. Scale bar, 30 µm. f The proportion of the βIII-tubulin+ cells that expressed Tbr1, Ctip2, Satb2, or Cux1 in cocultures with or without microglia (two-sided Mann–Whitney U test; n = 10 independent experiments; P = 0.0021 for Tbr1, P = 7.6 x 10−5 for Ctip2, P = 7.6 x 10−5 for Satb2, P = 1.5 x 10−4 for Cux1). g An experimental schematic of the coculture of youngest (0 day, Gadd45g-d4Venus+) neurons and CX3CR1-GFP+ microglia. h Representative immunostaining (shown in pseudo color) for βIII-tubulin (cyan), Tbr1/Satb2/Cux1 (red), and Ctip2 (green). The arrowhead indicates microglia. Scale bar, 30 µm. i The proportion of βIII-tubulin+ cells that expressed Tbr1, Ctip2, Satb2, or Cux1 in cocultures with or without microglia (two-sided Mann–Whitney U test; n = 10 independent cultures; P = 0.529 for Tbr1, P = 0.289 for Ctip2, P = 0.631 for Satb2, P = 0.912 for Cux1). Data are presented as the mean values ± S.D. Source data are provided as a Source Data File.

• IHC

CiteAb

Immunohistochemistry - Anti-TBR1 antibody (AB31940)

Immunohistochemistry-immunofluorescence using Anti-TBR1 antibody, ab31940. Publication image from Hattori, Y. et al., 2020, Nat Commun, 32242005. Legend direct from paper.

In vitro-prepared CP-like neurons changed subtype marker expression upon exposure to microglia.a Immunostaining for d4Venus (Gadd45g; green), Tbr2 (red), MAP2 (cyan), and DAPI (blue) in E14 Gadd45g-d4Venus Tg mouse cerebral walls. The broken lines indicate apical and basal surfaces. Scale bar, 50 µm. b The protocol for the in vitro preparation of CP-like neurons, which were obtained from Gadd45g-d4Venus+ cells isolated from the E14 Gadd45g-d4Venus cortex by FACS, after culture for 2 days. c A time-lapse series showing the reduction in Gadd45g-dVenus expression during culture. Scale bar, 20 µm. d CX3CR1-GFP+ microglia harvested from E15 CX3CR1-GFP pallial walls by FACS were added to the CP-like neuronal cultures. e Representative immunostaining (shown in pseudo color) for βIII-tubulin (cyan), Tbr1/Satb2/Cux1 (red), and Ctip2 (green). The arrowhead indicates microglia. Scale bar, 30 µm. f The proportion of the βIII-tubulin+ cells that expressed Tbr1, Ctip2, Satb2, or Cux1 in cocultures with or without microglia (two-sided Mann–Whitney U test; n = 10 independent experiments; P = 0.0021 for Tbr1, P = 7.6 x 10−5 for Ctip2, P = 7.6 x 10−5 for Satb2, P = 1.5 x 10−4 for Cux1). g An experimental schematic of the coculture of youngest (0 day, Gadd45g-d4Venus+) neurons and CX3CR1-GFP+ microglia. h Representative immunostaining (shown in pseudo color) for βIII-tubulin (cyan), Tbr1/Satb2/Cux1 (red), and Ctip2 (green). The arrowhead indicates microglia. Scale bar, 30 µm. i The proportion of βIII-tubulin+ cells that expressed Tbr1, Ctip2, Satb2, or Cux1 in cocultures with or without microglia (two-sided Mann–Whitney U test; n = 10 independent cultures; P = 0.529 for Tbr1, P = 0.289 for Ctip2, P = 0.631 for Satb2, P = 0.912 for Cux1). Data are presented as the mean values ± S.D. Source data are provided as a Source Data File.

• IHC

CiteAb

Immunohistochemistry - Anti-TBR1 antibody (AB31940)

Immunohistochemistry-immunofluorescence using Anti-TBR1 antibody, ab31940. Publication image from Hattori, Y. et al., 2020, Nat Commun, 32242005. Legend direct from paper.

Artificial delivery of microglia into the CP disturbed the expression of subtype-associated transcription factors in postmigratory neurons.a A schematic showing the strategy of microglial attraction into the CP in E15 CX3CR1-GFP cortical slices using CXCL12-soaked beads. b An image of a cortical slice on which CXCL12-soaked beads were placed. Scale bar, 200 µm. c The microglial trajectories (1-h intervals for 24 h) in cultured slices. d A graph depicting the proportion of microglia localized in the CP or the inner region (VZ/SVZ/IZ) during culture. e Immunostaining for GFP (CX3CR1; green) and DAPI (blue) of the slice 24 h after culturing showing that microglia were attracted to the CP. f Representative immunostaining (shown in pseudo color) for GFP (CX3CR1; red), Tbr1/Ctip2/Satb2/Cux1 (green), and DAPI (blue) in cultured slices on which empty or CXCL12 beads were placed and in control slices without beads. g The density of cells expressing each marker per total (DAPI+) cells in 20 µm bins in the CP, which was divided into six zones and numbered from the inside. h The experimental design for in vivo CXCL12 overexpression in postmigratory neurons. i Two adjacent sections of a representative E16 CXCL12-overexpressing brain. In situ hybridization for Cxc12 mRNA (top) and anti-RFP antibody immunostaining (for Lyn-TdTomato) (lower two panels) are shown. j Immunostaining for RFP (Lyn-TdTomato), GFP (CX3CR1), and DAPI. In Cont. group, only Lyn-TdTomato was overexpressed in the brain. In CXCL12 and CXCL12 with CL injection groups, CXCL12 was overexpressed together with Lyn-TdTomato with or without intraventricular injection of CL. k Representative immunostaining for Tbr1/Ctip2/Satb2/Cux1 and DAPI in the CP of control and CXCL12-overexpressing brains with or without clodronate administration. l The density of cells expressing each marker per total (DAPI+) cells in 20 µm bins in the CP, which was divided into eight zones. For g and l, two-sided Steel–Dwass test was applied (n = 8 sections from four mice; ***P < 0.001, **P < 0.01, *P < 0.05, or NS not significant). Data are presented as the mean values ± S.D. The exact P values are summarized in Supplementary Table 1. Except for in b, the scale bar indicates 50 µm. Source data are provided as Source Data File.