PA5-20007

antibody from Invitrogen Antibodies

Targeting: BBC3 JFY1, PUMA

Western blot

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Validation data

Product number: PA5-20007 - Provider product page
Provider: Invitrogen Antibodies
Product name: PUMA alpha Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: The PA5-20007 immunogen is located within the last 50 amino acids of PUMA. The immunogenic sequence is identical between the A and B forms of PUMA. In Western blot applications, this antibody detects a band at ~23kDa. A suggested positive control is K562 cell lysate. PA5-20007 can be used with blocking peptide PEP-0127. Predicted species reactivity based on immunogen sequence: Rat: (100%), Mouse: (100%)
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C

BBC3 protein structure - PA5-20007 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of K562 cells using a PUMA polyclonal antibody (Product # PA5-20007) at a 2 µg/mL dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry of K562 cells using PUMA Polyclonal Antibody (Product # PA5-20007) at 1 µg/mL. Cells were fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1:250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with PUMA Polyclonal Antibody (Product # PA5-20007) at 2 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (red). Image showing cytosol staining on K562 cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 4% paraformaldehyde-fixed K562 cells labeling PUMA with PUMA Polyclonal Antibody (Product # PA5-20007) at 2 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (red). Image showing cytosol staining on K562 cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry of K562 cells using PUMA Polyclonal Antibody (Product # PA5-20007) at 1 µg/mL. Cells were fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1:250 was used as secondary. Counter stained with Hematoxylin.

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 5 CircLDB2 regulated non-squamous NSCLC development and cisplatin sensitivity in vitro by miR-346. Vector-transduced or circLDB2-infected A549 and H460 cells were transfected with miR-NC mimic or miR-346 mimic. (a) and (b) CCK-8 assay of cell proliferation. (c) Representative images depicting a colony formation assay. (d) Representative images showing a cell apoptosis assay and flow cytometry for cell apoptosis. (e) Wound-healing assay of cell migration. (f) Cell invasion by transwell assay. (g) and (h) transfected cells were exposed to various concentrations of cisplatin, followed by the evaluation of cell viability by CCK-8 assay. (i) and (j) Western blot showing the levels of p53 and PUMA in transfected cells. * p < 0.05

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 7 LIMCH1 was a functional target of miR-346. (a) and (b) Western blot analysis of LIMCH1 protein level in cells transfected with sh-NC or sh-LIMCH1. A549 and H460 cells were transfected with anti-NC + sh-NC, anti-miR-346 + sh-NC or anti-miR-346 + sh-LIMCH1. (c) and (d) CCK-8 assay of proliferation of transfected cells. (e) Representative images depicting a colony formation assay. (f) Representative images depicting a cell apoptosis assay and cell apoptosis by flow cytometry. (g) Wound-healing assay of cell migration. (h) Transwell assay of cell invasion. (i) and (j) CCK-8 assay for viability of transfected cells under cisplatin exposure. (k) and (l) Western blot showing the levels of p53 and PUMA in transfected cells. * p < 0.05

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 8 Overexpression of circLDB2 promoted the antitumor effect of cisplatin in vivo. The xenograft tumors were generated by subcutaneous injection of vector-transduced or circLDB2-transduced A549 cells (2 x 10 6 per mouse, n = 5 per group), and cisplatin treatment was performed twice a week by intraperitoneal injection. Mice were terminated at day 28 after cell implantation. (a) Representative images of the xenograft tumors. (b) Growth curves of the excised tumors. (c) Tumor average weight of the xenograft tumors. (d) Western blot showing the levels of LIMCH1, p53, and PUMA in the excised tumors. * p < 0.05

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 3 CircLDB2 overexpression regulated cell proliferation, apoptosis, migration, invasion, and cisplatin sensitivity in vitro. A549 and H460 cells were transduced with negative control lentivirus (vector) or circLDB2 overexpression lentivirus (circLDB2). (a) and (b) CCK-8 assay of proliferation of transduced cells. (c) Representative images depicting a colony formation assay and colony formation assay of colony formation ability. (d) Representative images depicting a cell apoptosis assay and cell apoptosis by flow cytometry. (e) Representative images showing a cell migration assay and cell migration by wound-healing assay. (f) Representative images showing a cell invasion assay and cell invasion by transwell assay. (g) and (h) CCK-8 assay of viability of transduced cells under cisplatin (various doses) exposure. (i) and (j) representative images depicting a western blot assay and western blot showing the levels of p53 and PUMA in transduced cells. * p < 0.05