PA5-51021

antibody from Invitrogen Antibodies

Targeting: NDUFA4 CI-9k, COXFA4, MLRQ

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-51021

Provider

Invitrogen Antibodies

Product name

NDUFA4 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

NDUFA4 Polyclonal Antibody detects endogenous levels of NDUFA4 protein.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Fascin Controls Metastatic Colonization and Mitochondrial Oxidative Phosphorylation by Remodeling Mitochondrial Actin Filaments.

Lin S, Huang C, Gunda V, Sun J, Chellappan SP, Li Z, Izumi V, Fang B, Koomen J, Singh PK, Hao J, Yang S
Cell reports 2019 Sep 10;28(11):2824-2836.e8

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Ndufa4 in RAW264.7 cells. Samples were probed using Ndufa4 Polyclonal Antibody (Product # PA5-51021).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), THP-1 (Lane 2), HeLa (Lane 3), SW-480 (Lane 4) and tissue extracts of Mouse Skeletal Muscle (Lane 5), Rat Skeletal Muscle (Lane 6), Mouse Heart (Lane 7) and Rat Heart (Lane 8). The blot was probed with Anti- NDUFA4 Polyclonal Antibody (Product # PA5-51021, 1µg/ml) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 9.5 kDa band corresponding to NDUFA4 was detected across the cell lines and tissues tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NDUFA4 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with NDUFA4 Rabbit Polyclonal Antibody (Product # PA5-51021) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). Panel d represents the merged image showing mitochondrial localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4. Fascin and mtF-Actin Regulate the Biogenesis of Respiratory Complexes in NSCLC (A) Waterfall plot showing fold changes of 12 mitochondrial proteins differentially regulated in fascin KO H1650 cells. Fold changes (log 2 ) are presented as mean +- SEM of results from two independent samples. (B and C) Western blotting showing the decrease in respiration Complex I subunits in fascin KO H1650 (B) and H23 (C) cells. (D and E) Western blotting showing the increase in the levels of Complex I subunits in H23 cells overexpressing wild-type fascin (D) but not the S39E and 149-151A mutants (E). (F) shRNA knockdown of INF2-CAAX abrogated fascin-mediated increase in mt-ND1 and NDUFB8 in H1650 cells. (G and H) The effect of fascin (G) and INF2-CAAX (H) depletion on the levels of respiratory super complexes in H1650 cells, as determined by BN-PAGE and western blotting using anti-OXPHOS antibody cocktails (recognized all 5 complexes) and antibodies specific for each respiratory complex (Cx I-V). S. C. indicates respirasomes consisting of Complex I, III, and IV. Representative results from at least three independent experiments are shown.