PA5-22396

antibody from Invitrogen Antibodies

Targeting: PNPT1 DFNB70, old-35, OLD35, PNPase

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-22396

Provider

Invitrogen Antibodies

Product name

PNPase Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, Jurkat, Raji, mouse liver.

Concentration

1 mg/mL

References

Submitted references

Identification of LACTB2, a metallo-β-lactamase protein, as a human mitochondrial endoribonuclease.

Levy S, Allerston CK, Liveanu V, Habib MR, Gileadi O, Schuster G
Nucleic acids research 2016 Feb 29;44(4):1813-32

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using PNPase Polyclonal Antibody (Product # PA5-22396). Sample (30 µg of whole cell lysate). Lane A: JurKat . 7.5% SDS PAGE. PNPase Polyclonal Antibody (Product # PA5-22396) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using PNPase Polyclonal Antibody (Product # PA5-22396). Sample (50 µg of whole cell lysate). Lane A: Mouse liver . 7.5% SDS PAGE. PNPase Polyclonal Antibody (Product # PA5-22396) diluted at 1:1,000.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using PNPase (Product # PA5-22396) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. LACTB2 is a mitochondrial, soluble and monomeric protein. ( A )Mitochondria localization. HeLa cells were disrupted and the nuclear,cytoplasmic and mitochondria-containing fractions were separated and analyzedfor the localization of LACTB2, using an immunoblotting assay. Thenucleus-located protein, CPSF3L, was used as marker for nuclear proteins.Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as a cytosolic markerand cytochrome C (Cyt C) as a mitochondrial marker. ( B ) Immunoblotanalysis of mitochondrial extract following Proteinase K accessibility assay.TX-100, Triton X-100. PK, Proteinase K. PNPase is a mitochondrial proteinlocated mainly in the intermembrane space and a small amount in the matrix.HSP60 is a mitochondrial matrix protein. Tom20 is a mitochondrial outermembrane protein. ( C ) LACTB2 is a soluble protein. Immunoblottinganalysis of mitochondrial extract following alkaline sodium carbonate(Na 2 CO 3 ) extraction of soluble and peripheral membraneproteins. T, total extract. S, supernatant. P, pellet. Subunit 5 of the ATPsynthase (ATP5) was used as a marker for intrinsic membrane proteins and Cyt Cas a marker of soluble proteins. (D) Immunoblot analysis of the recombinantLACTB2 (Rec.) and the native protein in isolated mitochondria (Mit.) showingthe same size on SDS-PAGE. Therefore, the mitochondria targeting signal is notcleaved upon entering the mitochondria and remain an integral part of themature protein. ( E ) LACTB2 is present as a monomer

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. LACTB2 is a mitochondrial, soluble and monomeric protein. ( A ) Mitochondria localization. HeLa cells were disrupted and the nuclear, cytoplasmic and mitochondria-containing fractions were separated and analyzed for the localization of LACTB2, using an immunoblotting assay. The nucleus-located protein, CPSF3L, was used as marker for nuclear proteins. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as a cytosolic marker and cytochrome C (Cyt C) as a mitochondrial marker. ( B ) Immunoblot analysis of mitochondrial extract following Proteinase K accessibility assay. TX-100, Triton X-100. PK, Proteinase K. PNPase is a mitochondrial protein located mainly in the intermembrane space and a small amount in the matrix. HSP60 is a mitochondrial matrix protein. Tom20 is a mitochondrial outer membrane protein. ( C ) LACTB2 is a soluble protein. Immunoblotting analysis of mitochondrial extract following alkaline sodium carbonate (Na 2 CO 3 ) extraction of soluble and peripheral membrane proteins. T, total extract. S, supernatant. P, pellet. Subunit 5 of the ATP synthase (ATP5) was used as a marker for intrinsic membrane proteins and Cyt C as a marker of soluble proteins. (D) Immunoblot analysis of the recombinant LACTB2 (Rec.) and the native protein in isolated mitochondria (Mit.) showing the same size on SDS-PAGE. Therefore, the mitochondria targeting signal is not cleaved upon entering the mitochondria and remain an integral part of the mature protein. ( E ) LACTB2 is