Anti-Dlx5 antibody [EPR4488]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dlx5 antibody [EPR4488] (AB109737)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human brain tissue labelling Dlx5 with unpurified ab109737 at 1/100.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-Dlx5 antibody [EPR4488] (AB109737)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Dlx5 antibody [EPR4488] (ab109737) at 1/5000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

U87-MG cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 32 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dlx5 antibody [EPR4488] (AB109737)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human astrocytoma tissue labelling Dlx5 with purified ab109737 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-Dlx5 antibody [EPR4488] (AB109737)

Blocking buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Dlx5 antibody [EPR4488] (ab109737) at 1/1000 dilution

All lanes:

GST and His tagged recombinant Human Dlx5 protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 60 kDa

false

Exposure time: 3s

• WB

CiteAb

Western blot - Anti-Dlx5 antibody [EPR4488] (AB109737)

Dlx5 western blot using anti-Dlx5 antibody [EPR4488] ab109737. Publication image and figure legend from Twomey-Kozak, J., Desai, S., et al., 2020, Int J Mol Sci, PubMed 32650430.

ab109737 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109737 please see the product overview.

DLX5 knockdown results in reduced expression of hypertrophy/ossification-related genes and decreased apoptosis in BM-MSCs. Relative mRNA expression levels of (A) DLX5, early chondrogenesis markers (B) COL1 and (C) COL2, terminal hypertrophic differentiation and ossification-related markers (D) COL10, (E) BGLAP and (F) ALPL in DLX5 siRNA knockdown BM-MSCs vs Control-scrambled siRNA BM-MSCs. The mRNA expression results are displayed at the basal level (“Noninduced” black-colored bars in the graph) and then again after undergoing chondrogenesis (“Chondro-induced” white-colored bars in the graph) to confirm the retention of the genotype. N ≥ 3; *, p ≤ 0.05; **, p ≤ 0.01 relative to control BM-MSCs. (G) Caspase-3/7 immunofluorescent staining of DLX5 siRNA knockdown BM-MSCs vs. Control-scrambled siRNA BM-MSCs. Scale = 250 µm. (H) Western blot analysis of DLX5, cleaved caspase-3, and the respective actin loading controls in DLX5 siRNA knockdown BM-MSCs and Control-scrambled siRNA BM-MSCs. Bar graphs represent the mean densitometric intensities of bands, normalized to Actin loading control bands. Measurements were made using ImageJ software. N = 3; *, p ≤ 0.05; ***, p ≤ 0.005.

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