Mitochondria Antibodies

The center of cellular signaling and energetic balance

The center of cellular signaling and energetic balance

Mitochondrial markers

Citric Acid Cycle

Mitochondrial Respiratory Complexes

Mitochondrial Fission

Mitochondrial Fusion

Mitochondrial Mediated Apoptosis

Mitochondrial  Autophagy

Mitochondrial Translation

Mitochondrial Protein Import

Introduction

Mitochondria are important cellular organelles that maintain cellular  energy balance, contain key regulators of cell death processes, and play  a significant role in cellular oxidative stress generation and maintenance  of  calcium homeostasis.

Links to cancer, apoptosis, autophagy, and hypoxia have brought  mitochondria to the forefront of scientific studies in recent years.  Knowledge of the subcellular location of a protein may reveal  the potential role it plays in a variety of cellular processes.

Proteintech offers approximately all the antibodies needed for mitochondria research.

   
  
Blog post:
Mitochondria: Center of cellular signaling and energetic balance
Mitochondria Research Slideshow

Mitochondrial markers

Mitochondria are composed of the inner and outer membranes, the intermembrane space, the cristae, and the matrix, and they contain their own DNA separated from the nucleus. Knowledge of the subcellular location of a protein may reveal the potential role it plays in a variety of cellular processes. Colocalization with one of the organelle-specific markers listed here can confirm the subcellular location of a mitochondrial protein of interest.

Antibody Name             
AIF COX7A2L HADHA SLC25A20
AK2 COXIV HSP60 SMCR7L
ALDH1B1 COXIV MFF SURF1
ATP5A1 CPT1A MFN2 SYNJ2BP
ATP5F1 ECH1 Mitofilin TOM20
ATP5H ECHS1 NDUFV2 TOM40
BCS1L ETFA NLRX1 UQCRC1
CLPP F1S1 ATP50  
COX2 GCSH OXCT1  
COX5B GLUD2 PMPCB  

 

HSPD1
Mitochondrial matrix and inner mitochondrial membrane marker
Immunofluorescent analysis of Hela cells, using HSPD1 antibody at 1:50 dilution and Rhodamine-labeled goat anti-mouse IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).
AIFM1
Mitochondrial inter-membrane space marker
Immunofluorescent analysis of HepG2 cells using AIF antibody at dilution of 1:25 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L)
SCOT
Mitochondrial matrix marker
Immunofluorescent analysis of MCF-7 cells, using OXCT1 antibody at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).
TOMM40
Mitochondrial outer membrane marker
Immunofluorescent analysis of HepG2 cells using TOMM40 Antibody at dilution of 1:25 and Rhodamine-Goat anti-Rabbit IgG

Citric Acid Cycle

The citric acid cycle – also known as the tricarboxylic acid (TCA) cycle or the Krebs cycle – is a series of chemical reactions used by all aerobic organisms to generate energy through the oxidation of acetate derived from carbohydrates, fats, and proteins into carbon dioxide and chemical energy in the form of adenosine triphosphate (ATP). In addition, the cycle provides precursors of certain amino acids as well as the reducing agent NADH that is used in numerous other biochemical reactions. In eukaryotic cells, the citric acid cycle occurs in the matrix of the mitochondrion. The reactions of the cycle are carried out by 8 enzymes that completely oxidize acetate, in the form of acetyl-CoA, into water and two molecules of carbon dioxide.

Related antibodies

Antibody Name
Aconitase 2   IDH3A
Citrate synthase   MDH1
DLD   OGDH
Fumarase   OGDHL
IDH2   SdhA
Blog post:
Mitochondria are more than the powerhouse of the cell

Mitochondrial Respiratory Complexes

Cellular respiration is the process that releases energy from food and supplies energy for life processes. The mitochondrial respiratory chain is the final and most important step for cellular respiration and is located on the inner membrane of the mitochondrion and comprises four large trans-membrane protein complexes (respiratory chain Complexes I, II, III, and IV) (CI, CII, CIII, CIV) as well as ubiquinone between CI/II and III and cytochrome c between CIII and IV. The function of the mitochondrial respiratory chain is biological oxidation by transferring electrons from NADH and succinate to oxygen and then generating proton gradient across the inner membrane.

Mitochondrial Fission

Mitochondria are integral to cellular function and are responsible for energy production in eukaryotes, synthesis of metabolites, phospholipids, and heme, and maintenance of calcium homeostasis. Mitochondria are remarkably dynamic organelles undergoing frequent fusion and fission events. The opposing processes of fission and fusion maintain mitochondrial morphology and it is this equilibrium that ensures maintenance of mtDNA and metabolic mixing, bioenergetic functionality, and organelle number.

 MFF  SMCR7L
IF analysis of (10% Formaldehyde ) fixed HepG2 cells using MFF antibody at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (green). IF analysis of Hela cells, using SMCR7L antibody at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red)

Mitochondrial Fusion

Mitochondrial fusion occurs in three stages:

1. Docking of two mitochondria via their outer membranes.

2. Fusion of outer membranes

3. Fusion of inner membranes. 

The first two stages are mediated by the mitofusion proteins: Mfn1 and Mfn2 in mammals. The next stage – fusion of the inner mitochondrial membranes – is mediated by the dynamin-related GTPase OPA1

Mitochondrial Mediated Apoptosis

Apoptosis is the process of programmed cell death (PCD) that plays a central role in animal development and tissue homeostasis. There are two major apoptotic pathways known to date, initiated by either the mitochondria (the ‘intrinsic’ pathway) or the cell surface receptors (the ‘extrinsic’ pathway). Mitochondria-mediated apoptosis occurs in response to a wide range of death stimuli, including activation of tumor suppressor proteins (such as p53) and oncogenes (such as c-Myc), DNA damage, chemotherapeutic agents, serum starvation, and ultraviolet radiation.

 

This pathway is initiated within the cell and results in increased mitochondrial permeability leading to release of pro-apoptotic molecules Cytochrome c into the cytoplasm.

Mitochondrial  Autophagy

Mitochondrial autophagy is the process of selective removal of damaged mitochondria by autophagosomes and subsequent catabolism by lysosomes. One common mechanism is that mitochondrial depolarization results in PINK1 stabilization and Parkin recruitment to the mitochondria. Parkin can ubiquitinate Mitofusins 1 and 2 (MFN1 and 2), hexokinases, TOM complex components, FIS1, BAK, MIRO, as well as VDAC, which may either be degraded through the proteasome or serve as binding partners for p62. p62 may in turn act as an adaptor molecule through direct interaction with LC3 to recruit autophagosomal membranes to the mitochondria. Parkin can also interact with Ambra1, which in turn activates the PI3K complex around mitochondria to facilitate selective mitophagy.

Related antibodies

Antibody Name  
BAK p62
Beclin 1 Parkin
FIS1 PINK1
Mfn1 VDAC1
Mfn2 VDAC2
NDP52 VDAC3
OPTN  

Mitochondrial Translation

The mitochondria contain their own DNA separated from the nucleus. During the course of evolution, most of the mitochondrial protein-coding genes have been transferred to the nuclear genome. However, a few genes have been retained in the genome of the modern organelle.We now know that the mitochondrial genome (mtDNA), which is housed in the mitochondrial matrix, contains the blueprint for thirteen proteins and all the RNA molecules believed to be necessary and sufficient for intra-mitochondrial protein synthesis. All the other required components for intra-mitochondrial protein synthesis are imported from the cytosol after their synthesis in the cytoplasm. Central to the process of mitochondrial protein synthesis is the mitochondrial ribosome.

MRPS18B 
IHC of paraffin-embedded human breast cancer, using MRPS18B antibody (16139-1-AP) at dilution of 1:50 (40x objective).
MRPS27
IHC of paraffin-embedded human kidney, using MRPS27 antibody (17280-1-AP) at dilution of 1:100 (40x objective)

Mitochondrial Protein Import

Though mitochondria possess their own genome and translation machinery, only a small number of mitochondrial proteins, including a few core constituents of the respiratory chain complexes, are encoded by mtDNA and synthesized within the organelle. Thus, the vast majority of mitochondrial proteins are nuclear-encoded and have to be imported into the organelle.Upon synthesis on free ribosomes, mitochondrial precursor proteins reach the surface of the organelle in a process that is guided by cytosolic chaperones. Subsequently, they are imported by specialized protein import machineries and sorted to the designated submitochondrial destination in the outer membrane, inner membrane, intermembrane space, or matrix.

TOM20
IHC of paraffin-embedded human gliomas, using TOM20 antibody (11802-1-AP ) at a dilution of 1:100 (10x objective).
TIMM23
IHC of paraffin-embedded human ovary tumor, using TIMM23 antibody (11123-1-AP)
at a dilution of 1:50 (10x objective).
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Loading control antibodies
GAPDH Antibody mouse monoclonal GAPDH antibody WB analysis of HeLa cells
Catalog no.: 60004-1-Ig

GAPDH is commonly used as a protein loading control in western blot due to its consistently high expression in most cell types. This enzyme participates in several cellular events such as glycolysis, DNA repair, and apoptosis.

Proteintech monoclonal GAPDH antibodies are raised against a whole-protein antigen of human origin and have over 4,960 citations.

Beta Actin Antibody (KD/KO validated) WB analysis of Jurkat cells using using beta actin antibody (66009-1-Ig)
Catalog no.: 66009-1-Ig 

Beta-actin is usually used as a loading control due to its broad and consistent expression across all eukaryotic cell types and the fact that expression levels of this protein are not affected by most experimental treatments.

66009-1-Ig has been cited in over 2,460 publications and has wide species reactivity.