Anti-AKAP 95 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKAP 95 antibody (AB72196)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (left) and mouse teratoma (right) tissues labelling AKAP8 with ab72196 at 1/1000 (0.2µg/ml) and 1/200 (1µg/ml). Detection : DAB.

• IP

Unknown

Immunoprecipitation - Anti-AKAP 95 antibody (AB72196)

Immunoprecipitation of HeLa whole cell lysate (1 mg) using ab72196 at 3 µg/mg lysate. 20% of the immunoprecipitate was used for Western blot and bands were detected using ab72196 at 1 µg/ml. Bands were developed using chemiluminescence with an exposure time of 3 seconds.
Lane 2 represents a Control IgG.

All lanes:

Immunoprecipitation - Anti-AKAP 95 antibody (ab72196)

Predicted band size: 76 kDa

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• WB

Supplier Data

Western blot - Anti-AKAP 95 antibody (AB72196)

All lanes:

Western blot - Anti-AKAP 95 antibody (ab72196) at 0.04 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Lane 4:

293T whole cell lysate at 50 µg

Predicted band size: 76 kDa

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Exposure time: 3min

• WB

CiteAb

Western blot - Anti-AKAP 95 antibody (AB72196)

Western Blotting using Anti-AKAP 95 antibody, ab72196. Publication image from Hu, X. et al., 2020, Nat Commun, 31980632. Legend direct from paper.

Functional screening to identify AKAP8 as an hnRNPM-interacting protein.a A flow chart showing the experimental approaches to identify hnRNPM-interacting proteins. b qRT-PCR analysis of the CD44v8 splicing reporter minigene screening for the candidate splicing factors. Data were plotted as the Log2 transformed v8 exon inclusion versus skipping with mean ± s.d, n = 3. Incl : Inclusion. c Western blot analysis showing the interactions between hnRNPM and its candidate interacting proteins. A Flag-tagged hnRNPM cDNA was transfected into the 293 cells and immunoprecipitated with a Flag antibody with or without RNase treatment. Antibodies recognizing specific candidates were used for western blot analysis. d Kaplan–Meier plot analysis of breast cancer patient distal metastasis-free survival (GSE20685, n = 237) showing that higher levels of AKAP8 expression predict lower metastatic potential. P value was calculated by log-rank test. e Kaplan–Meier plot analysis of the METABRIC breast cancer data set (n = 1758) showing that higher expression of AKAP8 shows better patient survival probability. f Box and whiskers plots with jitters representing distribution of AKAP8 mRNA expression levels in luminal A (LumA), claudin low (CLOW), and basal (Basal) breast cancers patients from the breast cancer METABRIC data set. The line within each box represents the median. Upper and lower edges of each box represent 75th and 25th percentile, respectively. The whiskers represent the maximum and minimum values within 1.5x the interquartile range. P values were calculated by two sample z test in e, f. Source data are provided as a Source Data file.

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• WB

CiteAb

Western blot - Anti-AKAP 95 antibody (AB72196)

Western Blotting using Anti-AKAP 95 antibody, ab72196. Publication image from Hu, X. et al., 2020, Nat Commun, 31980632. Legend direct from paper.

Depletion of AKAP8 promotes an EMT phenotype.a Western blot analysis of EMT markers using lysates from HMLE/Twist-ER cells expressing control shRNA (Ctrl) or AKAP8 shRNAs (KD-1, KD-2). Lysates were collected before (untreated) and after 12 days of tamoxifen (TAM) induction. b Phase-contrast images (x 10) displaying cell morphology differences between control (Ctrl) and AKAP8-silenced HMLE/Twist-ER cells before (untreated) and after 12 days of TAM induction. White line represents scale bar at 100 µm. c Immunofluorescence images (x 40) showing the loss of E-cadherin at cell junction 12 days after TAM treatment in the AKAP8 knockdown cells. Green : E-cadherin, Blue : DAPI. d Phase-contrast images of HCT116 cells showing cell morphology changes upon AKAP8 knockdown. White line represents scale bar at 100 µm. e Western blot analysis of the epithelial markers in HCT116 cells expressing control or AKAP8 shRNA. Source data are provided as a Source Data file.

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• WB

CiteAb

Western blot - Anti-AKAP 95 antibody (AB72196)

Western Blotting using Anti-AKAP 95 antibody, ab72196. Publication image from Hu, X. et al., 2020, Nat Commun, 31980632. Legend direct from paper.

Depletion of AKAP8 promotes an EMT phenotype.a Western blot analysis of EMT markers using lysates from HMLE/Twist-ER cells expressing control shRNA (Ctrl) or AKAP8 shRNAs (KD-1, KD-2). Lysates were collected before (untreated) and after 12 days of tamoxifen (TAM) induction. b Phase-contrast images (x 10) displaying cell morphology differences between control (Ctrl) and AKAP8-silenced HMLE/Twist-ER cells before (untreated) and after 12 days of TAM induction. White line represents scale bar at 100 µm. c Immunofluorescence images (x 40) showing the loss of E-cadherin at cell junction 12 days after TAM treatment in the AKAP8 knockdown cells. Green : E-cadherin, Blue : DAPI. d Phase-contrast images of HCT116 cells showing cell morphology changes upon AKAP8 knockdown. White line represents scale bar at 100 µm. e Western blot analysis of the epithelial markers in HCT116 cells expressing control or AKAP8 shRNA. Source data are provided as a Source Data file.

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• WB

CiteAb

Western blot - Anti-AKAP 95 antibody (AB72196)

Western Blotting using Anti-AKAP 95 antibody, ab72196. Publication image from Hu, X. et al., 2020, Nat Commun, 31980632. Legend direct from paper.

AKAP8 suppresses breast cancer metastasis.a Western blot analysis showing the decreased expression of epithelial markers, E-cadherin, γ-catenin, and Occludin in HIM3 cells expressing control shRNA (Ctrl) and AKAP8 shRNAs (KD-1, KD-2). b Schematic of a xenograft model to measure breast cancer metastasis using the HIM3 PDX cells and the LM2 breasts cancer metastatic cells. c–f Tail vein injection of the HIM3 PDX tumor cells showing that silencing AKAP8 promotes metastatic tumor growth. Normalized bioluminescent imaging (BLI) signals c and representative BLI images d were shown at indicated time points. The areas of lung metastasis nodules were quantified by image J e and representative lung H&E stains f were displayed N = 5. e The middle line indicates mean of lung metastatic area, the top and bottom lines represent s.d. In f, black line represents scale bar at 2 mm. g–i Tail vein injection of LM2 cells showing that ectopic expression of empty control (Ctrl) and AKAP8 (AKAP8 OE) inhibits metastatic tumor growth. Normalized BLI signals g, representative BLI images h, lung H&E stains i, and quantifications of total lung metastasis areas j were shown (N > 7). i black line represents scale bar at 2 mm. j The middle line indicates mean of lung metastatic area, the top and bottom lines represent s.d. All Error bars indicate s.e.m. (*) P < 0.05; (**) P < 0.01. N > 7. P values were tested by Student’s t test, two-tailed in c, e, g, j. Source data are provided as a Source Data file.

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