Ab119684

Manufactured by Abcam

Sourced in United Kingdom

Ab119684 is a laboratory equipment product offered by Abcam. It is designed for use in scientific research applications. The core function of this product is to [factual description of the product's core function, without interpretation or extrapolation].

Automatically generated - may contain errors

Lab products found in correlation

6 protocols using ab119684

Immunoprecipitation and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Trypsinized human cells were washed three times with PBS, and homogenized with a Dounce homogenizer together with IP lysis buffer (50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 1 mM EDTA, 5% glycerol, 0.1% SDS and 1% NP-40). The lysates were incubated on ice for 20 min and centrifuged at 12,000 × g at 4°C for 10 min to remove cell debris. 2–4 mg of cell extracts were incubated overnight at 4°C with 2 μg of mouse IgG (A0919; Merck), TFAM (mouse, ab119684; Abcam), or MCM8 (mouse H00084515-M02; Novus Biologicals) antibody. Immunoprecipitates were collected with high-affinity protein G agarose beads (ab193258; Abcam) for 3 h at 4°C, centrifuged at 200 g at 4°C for 5 min, and washed five times with IP lysis buffer. Samples were eluted with Bolt LDS sample buffer for 5 min at 95°C, and separated on a Bolt 4–12% gel as described above.

Klucnika A., Mu P., Jezek J., McCormack M., Di Y., Bradshaw C.R, & Ma H. (2022). REC drives recombination to repair double-strand breaks in animal mtDNA. The Journal of Cell Biology, 222(1), e202201137.

+ Open protocol

+ Expand

Immunostaining of Stem Cell and Neuronal Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mounted sections were incubated for 1 h at room temperature with and blocked using blocking buffer containing (Sigma‐Aldrich, #G9023) and 0.1% (v/v) Triton X100 (Sigma‐Aldrich, #9036‐19‐5), and then incubated with primary antibodies diluted in blocking solution overnight at 4 °C. The following primary antibodies were used for immunostaining: Nanog (rabbit, 1:100; Abcam, #ab80892), OCT4 (rabbit, 1:100; Abcam, #ab19857), SSEA4 (mouse, 1:200; Abcam, #ab16287,), NeuN (rabbit, 1:500; Cell Signaling, #24307S), MAP2 (rabbit, 1:1000; Abcam, #ab5392), β‐tubulin III (TUJ1) (mouse, 1:1000; Abcam, #ab78078), Synaptophysin (mouse, 1:500; Proteintech, #17785‐1‐AP), PSD‐95 (rabbit, 1:500; Proteintech, #20665‐1‐AP), SOX2 (rabbit, 1:100; Abcam, #ab97959), NDUFB10 (rabbit, 1:350; Abcam 196 019), TFAM (mouse, 1:1000; Abcam #ab119684), TOMM20 (mouse, 1:350; Abcam, #ab56783), GFAP (chicken, 1:500; Abcam #ab4674), DCX (mouse, 1:200; Abcam #ab135349), SATB2 (rabbit, 1:400; Abcam #ab4674), CTIP2 (rat, 1:500; Abcam, #ab18465), OLIG2 (rabbit, 1:500; Abcam, #ab42453). Alexa Fluor Dyes (Invitrogen, #A11008, #A21449, #A11012, #A21141, #A11042, #A21236) were used at 1:800 dilution as secondary antibodies. Slides were mounted using ProLong diamond antifade mounting medium (SouthernBiotech, #0100‐20), and analyzed using the Leica TCS SP8 STED 3X (Leica microsystems).

Chen A., Yangzom T., Hong Y., Lundberg B.C., Sullivan G.J., Tzoulis C., Bindoff L.A, & Liang K.X. (2024). Hallmark Molecular and Pathological Features of POLG Disease are Recapitulated in Cerebral Organoids. Advanced Science, 11(18), 2307136.

+ Open protocol

+ Expand

Western Blot Analysis of Metabolic Regulators

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot was performed as previously described (22 (link)) by using the following antibodies: PGC-1α (P-19; sc-5815, 1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA), nuclear respiratory factor-1 (NRF-1; ab55744, 1:500; Abcam, Cambridge, United Kingdom), transcription factor A, mitochondrial (TFAM; ab119684, 1:500; Abcam), AMPK (2532, 1:1000; Cell Signaling Technology, Danvers, MA, USA), phospho-AMPK-α for the detection of endogenous AMPKα only when phosphorylated at threonine 172 (2531, 1:1000: Cell Signaling Technology), serine–threonine liver kinase B1 (LKB1; ab15095, 1:100; Abcam), and mammalian target of rapamycin (mTOR; ab87540, 1:200; Abcam).

Imasawa T., Obre E., Bellance N., Lavie J., Imasawa T., Rigothier C., Delmas Y., Combe C., Lacombe D., Benard G., Claverol S., Bonneu M, & Rossignol R. (2016). High glucose repatterns human podocyte energy metabolism during differentiation and diabetic nephropathy. The FASEB Journal, 31(1), 294-307.

+ Open protocol

+ Expand

Immunostaining of Mitochondrial Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

In brief, cells were fixed in 4% PFA (Sigma-Aldrich) for 15 min, followed by a 10-min permeabilization step (0.1% Triton X-100 in PBS for internal cell markers). The blocking step was performed by incubation in 2% BSA for 30 min. Cells were incubated with primary antibodies at 4°C overnight and further incubated with secondary antibodies for 1 h. For MitoTracker dye staining, cells were cultured in the normal media with 150 nM MitoTracker red (Invitrogen) for 30 min before further fixation and immunostaining. Antibodies against the following proteins were used at the indicated dilutions: CHCHD2 (HPA027407, 1:200; Sigma-Aldrich), SOX1 (4194S, 1:200; Cell Signaling Technology), NESTIN (MAB5326, 1:200; EMD Millipore), TUJ1 (MRB-435P, 1:500; Covance), mtTFA (ab119684, 1:500; Abcam), anti–mouse IgG-FITC (1:800; Sigma-Aldrich), and anti–rabbit IgG-Cy3 (1:1,000; Jackson ImmunoResearch Laboratories, Inc.). Nuclei were labeled with DAPI (Thermo Fisher Scientific). Colocalization coefficient studies were performed using ImageJ software by calculating Manders’ colocalization coefficient, which describes the amount of colocalizing pixels of GFP using pixels generated by RFP.

Zhu L., Gomez-Duran A., Saretzki G., Jin S., Tilgner K., Melguizo-Sanchis D., Anyfantis G., Al-Aama J., Vallier L., Chinnery P., Lako M, & Armstrong L. (2016). The mitochondrial protein CHCHD2 primes the differentiation potential of human induced pluripotent stem cells to neuroectodermal lineages. The Journal of Cell Biology, 215(2), 187-202.

+ Open protocol

+ Expand

Immunogold Labeling of Mitochondrial Transcription Factor A

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified human spermatozoa were collected and fixed with a freshly made fixative containing 3% paraformaldehyde and 0.1% glutaraldehyde in 0.1M phosphate buffer with 4% sucrose (pH 7.2). After washing with 0.1M phosphate buffer with 4% sucrose, 0.1M glycine was added to quench the unbonded aldehyde group. The cells were dehydrated in graded series of ethanol on ice and embedded in LR White (Electron Microscopy Sciences, Hatfield, PA). The samples were polymerized under UV light (360nm) at −10°C for 48 h, followed by 12 h at RT. The 90 nm-thin sections were cut and mounted on Formvar-Carbon coated 200 mesh nickel grids. The grid was incubated with an anti-TFAM antibody (1:1000, Abcam, #ab119684) in PBS with 1% BSA, and 0.05% Tween 20, for 2 h at RT, then overnight at 4°C. Upon washing, the anti-mouse secondary antibody conjugated with 18 nm gold (1:15, Colloidal Gold AffiniPure Goat Anti-Mouse IgG (H+L, EM Grade, Jackson ImmunoReasearch Laboratories, Inc.,) were added in PBS with 1% BSA, and 0.05% Tween 20, and incubated for 1h. After washing, the grids were stained with uranyl acetate and lead citrate by standard methods, imaged with Talos120C transmission electron microscope (Thermo Fisher Scientific), and recorded using Gatan (4k x 4k) OneView Camera with software Digital Micrograph (Gatan Inc).

Lee W., Zamudio-Ochoa A., Buchel G., Podlesniy P., Gutierrez N.M., Puigròs M., Calderon A., Tang H.Y., Li L., Mikhalchenko A., Koski A., Trullas R., Mitalipov S, & Temiakov D. (2023). Molecular Basis for Maternal Inheritance of Human Mitochondrial DNA. Nature genetics, 55(10), 1632-1639.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Mitochondrial Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The expression levels of specific proteins of the pathological slices were determined by IHC staining. After dewax, rehydration, peroxidase quenching and antigen-retrieval procedures, the pathological slices were subjected to reaction with primary antibodies (antigen retrieval reagent/reaction time, name of primary antibody/product name/company/dilution fold/reaction time) against TFAM (citrate/10 min, anti-mtTFA antibody/ab119684/Abcam/1:1600/30 min), [ 20 ] NADH dehydrogenase subunit 1 (ND1), (EDTA/10 min, anti-MT-ND1 antibody/ab218027/Abcam/1:600/30 min), [ 21 ] PDH (citrate/20 min, anti-PDH E1-alpha subunit phospho S293 antibody/ab92696/abcam/1:400/30 min) [ 22 ] and HK-II (EDTA/20 min, anti-HK-II antibody/ab37593/Abcam/1:400X/30 min), [ 23 ] followed by specific secondary antibodies and detection methods (polymer-HRP/DAB), respectively, according the literature and the manufacturer's instruction. The relative protein expression levels were scored semi-quantitatively by the H-score. [ 24 ] H-score = 3 × percentage of strongly stained cells + 2 × percentage of moderately stained cells + 1 × percentage of weakly stained cells, 0 ≤ H-score ≤ 300.

Lin C., Yeh Y., Pan S., Lu S., Chen Y., Chueh W., & Wei Y. (2020). Role of mitochondrial DNA copy number alteration in non-small cell lung cancer. Formosan Journal of Surgery, 53(5), 165-176.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!