Human/Mouse/Rat HTRA2/Omi Antibody

Detection of Human HTRA2/Omi by Western Blot

Caspases are not activated by FGFR inhibition in FGFR2‐mutant EC cells. (A) Western blots showing total caspase‐3 and caspase‐7 in response to treatment with 300 nm BGJ398 for up to 72 h. Tubulin serves as a loading control. *Denotes nonspecific band. (B) AN3CA and JHUEM2 cells were pretreated with 100 μm Z‐VAD‐FMK for 1 h prior to the addition of DMSO, 1 μm PD173074 (PD), 300 nm BGJ398 (BGJ) or 300 nm AZD4547 (AZD) for 72 h. Cell death was detected by staining cells with Annexin V. The mean percentage of Annexin V‐positive cells from three independent experiments (each performed in triplicate) is shown along with SD. (C) Western blot showing cleavage of caspase‐3 in AN3CA and JHUEM2 cells treated with 1 μm actinomycin D (Act D) or staurosporine (STS), respectively, for 24 h. (D) Mean percentage of AN3CA and JHUEM2 cells showing Annexin V‐positive staining following treatment with 100 μm Z‐VAD‐FMK alone or 1 h prior to treatment with 1 μm Act D and STS for 24 h. The mean from three independent experiments (each performed in triplicate) is shown along with SD. (E) Western blot showing staining of Bim, Bid, HTRA2/OMI and Smac/diablo in cytosolic and mitochondrial fractions of JHUEM2 cells treated with 300 nm BGJ398 for 24 and 48 h. Tom20 serves as a marker of the mitochondrial fraction, GRP78 as a marker of the ER, Lamin B1 as a marker of the nuclear fraction and GAPDH as a marker of the cytosolic fraction. (F) Western blot showing staining of AIF in mitochondrial and nuclear fractions of AN3CA cells treated with 1 μm PD173074 for 48 h. Cox IV and PARP serve as mitochondrial and nuclear markers, respectively. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30537101), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse HTRA2/Omi by Western Blot

Neural deletion of Htra2 is sufficient to generate neurological phenotypes.(A) Exons 2 to 4 of Htra2 were flanked with loxP sites, with a FRT flanked neo cassette 3′ to exon 4. Expression of FlpE causes deletion of the selection cassette. Cre-mediated deletion causes excision of exons 2 to 4. Small arrows beneath the allele constructs denote the position of genotyping primers. (B) PCR from genomic DNA can distinguish WT (+, arrow, 279 bp), KO (–, filled arrowhead, 358 bp) and floxed (f, empty arrowhead, 313 bp) alleles of Htra2. (C) Western blot analysis confirmed loss of HTRA2 protein (arrow) in all tissues of HTRA2 KO mice and reduction in brain of NesKO mice (arrowheads denote non-specific bands). The levels of HTRA2 protein in NesKO spleen and thymus were comparable with NesWT. Cx: cortex, Mb: midbrain, Hb: hindbrain. PHB2 was used as a loading control. (D) HTRA2 KO mice and NesKO mice were smaller than WT littermates by comparison. The size of the thymus and spleen was reduced although brain was relatively normal in size (representative animals shown at P30, scale bar: 1 cm.). (E) Body weight of HTRA2 KO and NesKO mice did not increase beyond P18 (n = 56 (HTRA2 WT), 62 (HTRA2 KO), 35 (NesWT), 25 (NesKO), error bars indicate SEM). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25531304), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse HTRA2/Omi by Western Blot

Neural deletion of Htra2 is sufficient to generate neurological phenotypes.(A) Exons 2 to 4 of Htra2 were flanked with loxP sites, with a FRT flanked neo cassette 3′ to exon 4. Expression of FlpE causes deletion of the selection cassette. Cre-mediated deletion causes excision of exons 2 to 4. Small arrows beneath the allele constructs denote the position of genotyping primers. (B) PCR from genomic DNA can distinguish WT (+, arrow, 279 bp), KO (–, filled arrowhead, 358 bp) and floxed (f, empty arrowhead, 313 bp) alleles of Htra2. (C) Western blot analysis confirmed loss of HTRA2 protein (arrow) in all tissues of HTRA2 KO mice and reduction in brain of NesKO mice (arrowheads denote non-specific bands). The levels of HTRA2 protein in NesKO spleen and thymus were comparable with NesWT. Cx: cortex, Mb: midbrain, Hb: hindbrain. PHB2 was used as a loading control. (D) HTRA2 KO mice and NesKO mice were smaller than WT littermates by comparison. The size of the thymus and spleen was reduced although brain was relatively normal in size (representative animals shown at P30, scale bar: 1 cm.). (E) Body weight of HTRA2 KO and NesKO mice did not increase beyond P18 (n = 56 (HTRA2 WT), 62 (HTRA2 KO), 35 (NesWT), 25 (NesKO), error bars indicate SEM). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25531304), licensed under a CC-BY license. Not internally tested by R&D Systems.