PA5-32043

antibody from Invitrogen Antibodies

Targeting: LACTB2 CGI-83

Provider product page for PA5-32043

Western blot

Immunohistochemistry

Other assay

Antibody data

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

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Product number: PA5-32043 - Provider product page
Provider: Invitrogen Antibodies
Product name: LACTB2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: A549, HCT116. Predicted reactivity: Mouse (82%), Rat (81%), Pig (86%), Bovine (89%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.61 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

LACTB2 protein structure - PA5-32043 shown in red.

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8. Downregulation of LACTB2 expression resulted in modest accumulation of mitochondrial transcripts. ( A ) Quantitation of the amount of LACTB2 in the siRNA treated cells. siRNA duplex targeted for LACTB2 or scrambled siRNAs duplexes, as a negative control, were transfected into HEK-293 cells. Cells were collected 48 h post-transfection and the amount of LACTB2 was determined using immunoblotting assay. Proteins from equal number of cells were loaded in the lanes labeled 100%. Proteins from 50, 25 and 10% of the control and 150 and 200% of the siRNA LACTB2 treated cells were analyzed for the amount of LACTB2, in order to determine the degree of down-expression. ( B ) Medium acidification due to treatment with siRNA duplex, 48 h post-transfection. Ctrl--cells treated with scrambled siRNAs duplexes as a negative control. RNAi--cells treated with siRNA duplex targeted to LACTB2. ( C ) The amount of mitochondrial transcripts in the cells treated with the siRNA duplex targeted to LACTB2 (turquoise bars), or with scrambled siRNAs duplexes as a negative control (brown bars), was analyzed by qRT-PCR. The GAPDH transcript was used as the reference gene for normalization of the expression. The error bars display the standard errors of at least three independent experiments. ( D ) Northern blot analysis of HEK-293 cells that were not transfected (WT), transfected with scrambled siRNAs as control (Ctrl) or transfected with LACTB2 siRNA (RNAi). Hybridizations were performed with pr