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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- 701696 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- BNIP3 Recombinant Rabbit Monoclonal Antibody (8H14L2)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Bovine and Equine.
- Antibody clone number
- 8H14L2
- Concentration
- 0.5 mg/mL
Submitted references Quantitative Ultrastructural Morphometry and Gene Expression of mTOR-Related Mitochondriogenesis within Glioblastoma Cells.
Ferese R, Lenzi P, Fulceri F, Biagioni F, Fabrizi C, Gambardella S, Familiari P, Frati A, Limanaqi F, Fornai F
International journal of molecular sciences 2020 Jun 27;21(13)
International journal of molecular sciences 2020 Jun 27;21(13)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hela (Lane1), Hela treated with Cobalt Chloride (15 µm/ 48 hours) (Lane 2), PC-3 (Lane 3) and PC-3 treated with Etoposide (25 uM/6hours) (Lane 4). The blots were probed with Anti-BNIP3 Recombinant Rabbit Monoclonal Antibody (Product # 701696, 0.5-1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 22 kDa band corresponding to BNIP3 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence was performed on methanol fixed Hela cells treated with Cobalt Chloride (15 µm/ 48 hours), for detection of BNIP3 using Anti-BNIP3 Recombinant Rabbit Monoclonal Antibody (Product # 701696, 1-2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of BNIP3 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of BNIP3. Panel e) represents merged image of untreated cells with no signal. Panel f) represents control cells with no primary Antibody to assess background.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 18 Rapamycin increases the mitophagy marker BNIP3 at short-time intervals within U87MG cells. ( A ) Representative BNIP3-stained mitochondrion (indicated by the black arrow) at 24 h during 10 nM rapamycin, continuous exposure. Graphs report: ( B ) the amount of total BNIP3 particles; ( C ) the number of BNIP3-positive mitochondria; ( D ) the ratio of mitochondrial/cytosolic BNIP3 particles at various time interval during (12-24 h) or following (from 1 d up to 14 d) rapamycin exposure. M = mitochondrion, AV = vacuoles. * p
