- PA5-22252 - Invitrogen Antibodies CASP9 antibody

Submitted references E2F5 Promotes the Malignancy of Ovarian Cancer Via the Regulation of Hippo and Wnt Pathways.

Malgundkar SH, Burney I, Al Moundhri M, Al Kalbani M, Lakhtakia R, Okamoto A, Tamimi Y
Genetic testing and molecular biomarkers 2021 Mar;25(3):179-186

Anti-Apoptotic Effect of Apelin in Human Placenta: Studies on BeWo Cells and Villous Explants from Third-Trimester Human Pregnancy.

Mlyczyńska E, Myszka M, Kurowska P, Dawid M, Milewicz T, Bałajewicz-Nowak M, Kowalczyk P, Rak A
International journal of molecular sciences 2021 Mar 9;22(5)

FAT4 silencing promotes epithelial-to-mesenchymal transition and invasion via regulation of YAP and β-catenin activity in ovarian cancer.

Malgundkar SH, Burney I, Al Moundhri M, Al Kalbani M, Lakhtakia R, Okamoto A, Tamimi Y
BMC cancer 2020 May 4;20(1):374

In Vitro Effects of Vaspin on Porcine Granulosa Cell Proliferation, Cell Cycle Progression, and Apoptosis by Activation of GRP78 Receptor and Several Kinase Signaling Pathways Including MAP3/1, AKT, and STAT3.

Kurowska P, Mlyczyńska E, Dawid M, Opydo-Chanek M, Dupont J, Rak A
International journal of molecular sciences 2019 Nov 19;20(22)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Caspase 9 using 30 µg of 293T lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a Caspase 9 polyclonal antibody (Product # PA5-22252) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Caspase 9 was achieved by transfecting HeLa cells with Caspase 9 specific siRNAs (Silencer® select Product # s2429 & s2430). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the Caspase 9 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-Caspase 9 Polyclonal Antibody (Product # PA5-22252, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Caspase 9.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2) and A549 (Lane 3). The blot was probed with Anti-Caspase 9 Polyclonal Antibody (Product # PA5-22252, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 46 kDa band corresponding to Caspase 9 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Caspase 9 was performed by separating 30 µg of untreated (–) and treated (+) HeLa whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a Caspase 9 Polyclonal Antibody (Product # PA5-22252) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Caspase 9 Polyclonal Antibody (Product # PA5-22252). Untreated (–) and treated (+) HeLa whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with Caspase 9 Polyclonal Antibody (Product # PA5-22252) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Caspase 9 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Caspase 9 Rabbit Polyclonal Antibody(Product # PA5-22252) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nucleus and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry (Paraffin) analysis of Caspase 9 was performed in paraffin-embedded human lung cancer tissue using Caspase 9 Polyclonal Antibody (Product # PA5-22252) at a dilution of 1:400.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Caspase 9 Polyclonal Antibody detects Caspase 9 protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded human lung cancer. Caspase 9 stained by Caspase 9 Polyclonal Antibody (Product # PA5-22252) diluted at 1:1,000. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Regulatory effects of FAT4 on the expression of proteins involved in EMT, Hippo, Wnt-beta-catenin, apoptotic, and retinoblastoma pathways by Western blotting. a . Relative expression variation of proteins. The expression of Vimentin ( p = 0.0001), YAP ( p = 0.0018), beta-catenin ( p = 0.001), Bcl2 ( p = 0.0001), cyclin D1 ( p = 0.0017) and cdk4 ( p = 0.0025) was higher in FAT4 siRNA treated cells as compared to control. beta-actin was used as an internal control. b . Western blot demonstrating bands for protein expression in control and FAT4 knocked cells. Full-length blots are presented in Supplementary figure S 1 . c . The ratio of phosphorylated to total YAP, GSK-3beta, beta-catenin, and retinoblastoma proteins following FAT4 repression. The pYAP/ YAP ratio was lower in FAT4 siRNA treated cells as compared to the control ( p = 0.0286). Similarly, pGSK-3-beta/GSK-3-beta ratio, and pbeta-catenin/beta-catenin ratio was lower in FAT4 siRNA treated cells ( p = 0.018, and p = 0.001 respectively) as compared to control. There was no significant difference in pRb/Rb ratio between FAT4 siRNA treated cells and control. MCAS cells treated with scrambled siRNA was used as control. Data represent mean and standard deviation from at least three independent experiments performed in triplicates

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Effect of apelin on protein expression of apoptotic factors in BeWo cells. The cells were incubated with apelin at doses 2 of (AP2) and 20 (AP20) ng/mL for 24, 48 and 72 h, and subsequently, Western blot analysis was performed to examine the expression of BCL2 (B-cell like lymphoma 2), BAX (Bcl-2-like protein 4), caspase 3, 8 and 9 and p53. Results are shown as stripes on gel image ( A ) and densitometry analysis relative to beta-actin ( B - F ). Experiments were independently performed and repeated three times ( n = 3). The data are arranged as means +- SEM. Different letters indicate significant differences ( p < 0.05) among groups; Control ( C ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Effect of vaspin on mRNA and protein expression levels of caspase-3, -8, and -9; BAX (bcl-2-like protein 4), and BCL2 (B-cell lymphoma 2). The cells were treated with 1 and 10 ng/mL vaspin for 24 h, and then, real-time PCR and Western blot analysis were performed to determine levels of caspase-3, -8, and -9 ( A , B ); BCL2 and BAX ( C , D ); and p53 ( E , F ) (full gel images are available in the Supplementary Materials ). Gene expression levels were normalised to glyceraldehyde 3-phosphate dehydrogenase ( GAPDH) , while proteins levels were normalized to actin. Experiments were independently performed and repeated five times ( n = 5). The data are plotted as the means +- SEM. Significance between control and vaspin treatments is indicated by * p < 0.05, ** p < 0.01 and *** p < 0.001; Control (C).

PA5-22252

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