701696

antibody from Invitrogen Antibodies

Targeting: BNIP3 Nip3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

701696

Provider

Invitrogen Antibodies

Product name

BNIP3 Recombinant Rabbit Monoclonal Antibody (8H14L2)

Antibody type

Monoclonal

Antigen

Other

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

8H14L2

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

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)

Western blot

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).

Immunocytochemistry

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.

Other assay

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