PA5-25406

antibody from Invitrogen Antibodies

Targeting: MTCH2 SLC25A50

Provider product page for PA5-25406

Western blot

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Other assay [2]

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Product number: PA5-25406 - Provider product page
Provider: Invitrogen Antibodies
Product name: MTCH2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 400 µL
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

MTCH2 protein structure - PA5-25406 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. Detection of endogenous MTCH2x. A, Western blotting performed using an antibody (anti-MTCH2x) generated against the peptide encoded by the ISR1 (RCGAGTVTFL) in HEK293 cell lysate. B, Western blotting performed using the same antibody in HEK293 cells transfected with MTCH2 -specific shRNAs. C, detection of exogenous overexpressed MTCH2x by anti-MTCH2x antibody in HEK293 cells. D, amount of MTCH2x relative to MTCH2 was estimated by performing a prolonged SDS-PAGE followed by Western blotting using anti-MTCH2 antibody that can potentially recognize all isoforms. Quantification is shown below (mean +- S.D., n = 3). Lysates from cells overexpressing MTCH2 or MTCH2x (without any tag) were used to distinguish endogenous MTCH2 and MTCH2x. E, detection of endogenous MTCH2x in mitochondrial extracts from multiple mouse organs.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8. Double-SCR of MTCH2 is required to maintain normal mitochondrial membrane potential. A, CRISPR-Cas9 technique was used to delete the most part of ISR1 and ISR2 in the genome of HEK293 cells (termed DeltaRT MTCH2 ). Deletion was confirmed by PCR amplification from genomic DNA using primers flanking the targeted region, and by sequencing the product. Sequencing was performed using a reverse primer that generated reverse-complement sequence. A part of this is shown as an electropherogram. The deleted part in ISR1 and ISR2 is shown in gray background. Three in-frame stop codons are shown in boldface . B, Western blotting showing the absence of MTCH2x and increased levels of MTCH2 in DeltaRT MTCH2 cells compared with the WT cells ( WT ). Long and short exposures are shown for MTCH2. C, JC-1 fluorescence profile shows reduced mitochondrial membrane potential in two different DeltaRT MTCH2 clones ( left ). H 2 O 2 treatment was used as a positive control for mitochondrial membrane depolarization. Ratio of fluorescence at lambda 594 to lambda 538 is shown ( right ) ( n = 4). D, DeltaRT MTCH2 cells show reduced ATP levels compared with the WT cells. Cells were incubated in serum-free DMEM containing 10 mu m galactose for 48 h before measuring ATP levels using a luminescence-based assay ( n = 3). E, WT and DeltaRT MTCH2 cells have comparable mitochondrial mass. The flow cytometry was performed using the fluorescent probe, nonyl acridine orange ( NAO ) ( n = 3). F, quantitativ