Mitochondrial Markers Antibody Pack

Western blot shows lysates of A431 human epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human COX4 Monoclonal Antibody (Catalog # MAB6980) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For addtional reference, recombinant human COX4-I1 (5 ng/lane) and recombinant human COX4-I2 (2.5 ng/lane) were included. A specific band was detected for COX4 at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Human HeLa cell line was stained with Mouse Anti- Cytochrome C Monoclonal Antibody (Catalog # NB100-56503, filled histogram), or Mouse IgG2B isotype control (Catalog # MAB0041, open histogram) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).Images may not be copied, printed or otherwise disseminated without express written permission of Novus Biologicals a bio-techne brand.

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with HSP60 Antibody (NBP1-77397) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Prohibitin was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Human Prohibitin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3470) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Total protein from Human HeLa cells, Mouse MEF cells and Rat Brain was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-OPA1 in 1% blocking buffer. Precision Plus Protein All Blue molecular weight markers (BioRad) were detected with 1 ug/ml Anti-Blue Marker antibody (NBP2-33376). The protein ladder was detected with an anti-mouse HRP secondary antibody and OPA1 with an anti-rabbit HRP secondary antibody using chemiluminescence.

Mitofusin-1 was detected in mouse liver by Immunohistochemistry using Mouse Anti-Mouse Mitofusin-1 (11E9-1H12) Monoclonal Antibody (NBP1-71775).

HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-Mitofusin 2 (NBP2-66383) at 5 ug/mL overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at 1:500. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at 1:1000 and detected with an anti-mouse DyLight 550 (Red) at 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC

Mitochondrial Markers Antibody Pack Summary

• Description: This pack contains 1 vial each of: MAB6980-SP (25 ug), MAB6980-SP (0.025 mg), NB100-56503SS (0.025 ml), NBP1-77397SS (25 ug), AF3470-SP (0.025 ml), NB110-55290SS (0.025 ml), NBP1-71775SS (0.025 mg), NBP2-24915SS (0.025 mg), NBP2-66383SS (0.5 mL), NL007 (0.5 mL), NL001 (0.5 mL).
• Immunogen: See individual datasheets.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Western Blot
• Application Notes: See individual datasheets of components for their validated applications

Reactivity Notes

See individual datasheets of components for their validated species

Packaging, Storage & Formulations

• Storage: Storage is content dependent.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Antibody Packs are guaranteed for 1 year from date of receipt.