PA5-79052

antibody from Invitrogen Antibodies

Targeting: CLPX

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-79052

Provider

Invitrogen Antibodies

Product name

CLPX Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

500 µg/mL

Storage

-20°C

References

Submitted references

Inactivity of Peptidase ClpP Causes Primary Accumulation of Mitochondrial Disaggregase ClpX with Its Interacting Nucleoid Proteins, and of mtDNA.

Key J, Torres-Odio S, Bach NC, Gispert S, Koepf G, Reichlmeir M, West AP, Prokisch H, Freisinger P, Newman WG, Shalev S, Sieber SA, Wittig I, Auburger G
Cells 2021 Nov 29;10(12)

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry analysis of CLPX using anti-CLPX antibody (Product # PA5-79052) . CLPX was detected in a section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-CLPX antibody (Product # PA5-79052) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry of CLPX in HeLa cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with CLPX Polyclonal Antibody (Product # PA5-79052) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 ( A ) Analysis of both PRLTS3 patients' (in triplicates) versus 7 healthy controls' skin fibroblast proteome profiles in volcano plots, highlighting the accumulated proteins in mitochondria. The significance threshold at 1.3 on the Y -axis corresponds to the p -value 0.05, and the X -axis shows the traditional cut-off at 1.5-fold effects, although genetic evidence has demonstrated a neurodegenerative process at old age to be triggered by a lower gain-of-function, such as 1.3-fold dosage of alpha-synuclein. ( B ) Analysis of the differences between both patients in a STRING interaction diagram, focused on mitochondrial proteins with significant accumulation in ClpP-mutant patient skin fibroblast global proteome, which showed 1.2-fold stronger change in the severely affected patient 58955 than in the milder patient 0006. ( C ) Comparison of all accumulations with nominal significance in both PRLTS3 fibroblasts that showed consistency with nominal significant accumulations in ClpP- MEFs, using a Venn diagram. A total of 23 effects were consistent between species, among which all mitochondrial proteins are listed below, employing their gene symbol. ( D ) Analyses of protein abundance in fibroblasts from a human control (red128) and the patient with stronger fold changes (58955), controlling the subcellular fractionation purity and loading by GAPDH (cytosolic marker), HSP60 (mitochondrial), or LAMIN A/C (nuclear) abundance. ( E ) Quantitative immunoblots for CLPX and