Anti-Protein CASP antibody [2A10] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Protein CASP (green) using ab54583 at 10 μg/ml in ICC/IF.

This image was generated using the ascites version of the product.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Formalin-fixed, paraffin-embedded human spleen tissue stained for Protein CASP with ab54583 at 5 μg/ml in immunohistochemical analysis.

This image was generated using the ascites version of the product.

• Flow Cyt

Unknown

Flow Cytometry - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Overlay histogram showing MCF7 (human breast adenocarcinoma cell line) cells stained with ab54583 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3 M glycine to block non-specific protein-protein interactions followed by the antibody (ab54583, 1 μg/1 x 10⁶ cells) for 30 minutes at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 μg/1 x 10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

This image was generated using the ascites version of the product.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Formalin-fixed, paraffin-embedded human malignant lymphoma, diffuse large B tissue stained for Protein CASP with ab54583 at 5 μg/ml in immunohistochemical analysis.

This image was generated using the ascites version of the product.

• sELISA

Supplier Data

Sandwich ELISA - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Detection limit of ab54583 is approximately 0.03 ng/ml as a capture antibody.

This image was generated using the ascites version of the product.

• WB

Supplier Data

Western blot - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Protein CASP antibody [2A10] - BSA and Azide free (ab54583) at 5 µg/mL

Lane 1:

Protein CASP transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

Lane 2:

Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate

Predicted band size: 77 kDa

false

• WB

Unknown

Western blot - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.

Expected MW 38 kDa.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Protein CASP antibody [2A10] - BSA and Azide free (ab54583) at 5 µg/mL

All lanes:

Tagged recombinant Protein CASP protein

Predicted band size: 114 kDa,167 kDa,185 kDa,217 kDa,22 kDa,41 kDa,629 kDa,70 kDa,71 kDa,77 kDa,83 kDa

false

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Immunocytochemistry-immunofluorescence using Anti-Protein CASP antibody [2A10] - BSA and Azide free, ab54583. Publication image from Meissner, A. et al., 2015, Nat Commun, 25799239. Legend direct from paper.

Consecutive isolation of Notch active progenitors recapitulates cortical lamination and glial fates.(a) Combined HES5 : : eGFP reporter expression and immunostainings of cortical layer specific neuronal markers : Early born neurons expressing TBR1, RELN and CTIP2 (top two panels), and late derived neurons expressing SATB2, POU3F2 and CUX1 (bottom two panels), are shown for NE, M-RG and L-RG progenitors that were subjected to neuronal differentiation. Insets for RELN/TBR1 and SATB2/POU3F2 show magnified areas within the image. Inset for CTIP/TUJ1 shows same magnification but a different view of neuronal axons. Images of HES5+ derived neurons are shown. Scale bars : 50 µm for images, 25 µm for Insets. Images of HES5− derived neurons and percentages of all cortical subtypes derived from both HES5+ and HES5− cells are presented in Supplementary Fig. 5. (b) Distribution of relative transcript abundance based on qPCR for selected stage-specific marker gene groups for either deep or upper layer neuronal progeny. Contributions of HES5+ and HES5− populations per each respective stage are shown. Marker gene groups for each progenitor stage were created by collapsing the normalized values of TBR1/RELN, CTIP2/FEZF2 and CUX1/CUX2/SATB2 (see Methods for details). Individual qPCR analyses for all genes tested at all stages are shown in Supplementary Fig. 4. (c) Same as in b. Here, cumulative neuronal marker levels based on relative transcript levels are shown (top). Note the decrease in total neuronal progeny shown in the lower panel, as the glial marker GFAP is upregulated in panel e. (d) Distribution of relative transcript abundance based on qPCR for selected stage-specific marker genes for indicated progenitor or neuronal cell markers. Contributions of HES5+ and HES5− populations per each respective stage from either untreated or DAPT treated cells are shown. Expression levels relative to HPRT of all four conditions (colour coded) were summed per each gene and plotted as a single bar. (e) Top : Combined HES5 : : eGFP reporter expression and immunostaining of the glial marker GFAP following differentiation of the L-RG stage. Scale bar : 50 µm. Bottom : GFAP transcript level for distinct progenitor stages as assessed by qPCR. Values were obtained from three technical replicates. Statistical analysis : mean±s.e.m.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Immunocytochemistry-immunofluorescence using Anti-Protein CASP antibody [2A10] - BSA and Azide free, ab54583. Publication image from Meissner, A. et al., 2015, Nat Commun, 25799239. Legend direct from paper.

Transition through progenitor cell stages demarcates developing rosettes as VZ and SVZ equivalents.(a) Differential expression levels for selected genes that are most differentially expressed between HES5+ and HES5− cells in a stage-specific manner. Selected gene members are indicated on the left, developmental stages are indicated on the bottom, and gene categories classified by stage are indicated on the right. Values plotted on the heatmap represent ratios of expression levels relative to ES cells. (b) Relative expression levels (z-scores) based on microarray expression data for the entire differentiation time course for selected germinal zone marker genes. Relative expression levels are shown for HES5+ (top) and HES5− (bottom) samples separately. Genes are ordered from VZ to SVZ and from neurogenic to gliogenic markers. Individual qPCR analyses for all genes tested at all stages are shown in Supplementary Fig. 6c. Note that the apparently high GFAP expression in HES5+ cells at the L-RG stage has in fact low absolute expression values, and only appear high relatively to expression in other stages (all stages per each gene are normalized to 1; that is, highest red intensity). To compare GFAP transcript levels during proliferation and serum induced astrocytic differentiation, see Figs 5d and 3e, respectively. (c) Combined HES5 : : eGFP reporter expression and immunostainings of neural stem/progenitor markers, RG markers, and proliferation markers throughout the progression period. From top : PAX6 marking the VZ and TBR2 marking the SVZ are shown. Middle : CUX1 marking SVZ is shown. Bottom : the (mainly) SVZ marker POU3F2 is shown. Scale bar : 50 µm (valid for all images in c). (d) High-power magnification of E-RG and M-RG images shown in c. Dashed lines demarcate proposed VZ, SVZ and OSVZ regions, containing apical RG, INPs and basal RG, respectively. Scale bar : 25 µm (valid for all images in d).

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Protein CASP antibody [2A10] - BSA and Azide free (AB54583)

Immunocytochemistry-immunofluorescence using Anti-Protein CASP antibody [2A10] - BSA and Azide free, ab54583. Publication image from Meissner, A. et al., 2015, Nat Commun, 25799239. Legend direct from paper.

Transition through progenitor cell stages demarcates developing rosettes as VZ and SVZ equivalents.(a) Differential expression levels for selected genes that are most differentially expressed between HES5+ and HES5− cells in a stage-specific manner. Selected gene members are indicated on the left, developmental stages are indicated on the bottom, and gene categories classified by stage are indicated on the right. Values plotted on the heatmap represent ratios of expression levels relative to ES cells. (b) Relative expression levels (z-scores) based on microarray expression data for the entire differentiation time course for selected germinal zone marker genes. Relative expression levels are shown for HES5+ (top) and HES5− (bottom) samples separately. Genes are ordered from VZ to SVZ and from neurogenic to gliogenic markers. Individual qPCR analyses for all genes tested at all stages are shown in Supplementary Fig. 6c. Note that the apparently high GFAP expression in HES5+ cells at the L-RG stage has in fact low absolute expression values, and only appear high relatively to expression in other stages (all stages per each gene are normalized to 1; that is, highest red intensity). To compare GFAP transcript levels during proliferation and serum induced astrocytic differentiation, see Figs 5d and 3e, respectively. (c) Combined HES5 : : eGFP reporter expression and immunostainings of neural stem/progenitor markers, RG markers, and proliferation markers throughout the progression period. From top : PAX6 marking the VZ and TBR2 marking the SVZ are shown. Middle : CUX1 marking SVZ is shown. Bottom : the (mainly) SVZ marker POU3F2 is shown. Scale bar : 50 µm (valid for all images in c). (d) High-power magnification of E-RG and M-RG images shown in c. Dashed lines demarcate proposed VZ, SVZ and OSVZ regions, containing apical RG, INPs and basal RG, respectively. Scale bar : 25 µm (valid for all images in d).