PA5-19471

antibody from Invitrogen Antibodies

Targeting: COX4I1 COX4, COX4-1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-19471

Provider

Invitrogen Antibodies

Product name

COX4 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Heat mediated antigen retrieval recommended prior to tissue staining. This antibody is predicted to react with chimpanzee based on sequence homology. Store antibody at 4ºC for 1-2 weeks. For long-term storage, store at -20ºC.

Reactivity

Human, Mouse, Rat, Hamster, Xenopus

Host

Rabbit

Isotype

IgG

Vial size

100 µg

Concentration

0.9 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

miR-378a-3p Participates in Metformin's Mechanism of Action on C2C12 Cells under Hyperglycemia.

Machado IF, Teodoro JS, Castela AC, Palmeira CM, Rolo AP
International journal of molecular sciences 2021 Jan 7;22(2)

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The Soluble Adenylyl Cyclase Inhibitor LRE1 Prevents Hepatic Ischemia/Reperfusion Damage Through Improvement of Mitochondrial Function.

Teodoro JS, Amorim JA, Machado IF, Castela AC, Steegborn C, Sinclair DA, Rolo AP, Palmeira CM
International journal of molecular sciences 2020 Jul 11;21(14)

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Mouse Skeletal Muscle Tissue Lysate using Product # PA5-19471, COX IV primary antibody at a dilution of 1 µg/mL (lane 1). Staining of Human Skeletal Muscle Tissue Lysate at a dilution of 1 µg/mL (lane 2). Blot treated with a secondary HRP-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:3000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Heart (Lane 1), Mouse Liver (Lane 2) and Mouse Brain (Lane 3). The blot was probed with COX4 Polyclonal Antibody (Product # PA5-19471, 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 band at 15kDa corresponding to COX4 was observed across the tissues tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of COX4 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with COX4 Polyclonal Antibody (Product # PA5-19471) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). 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.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical (formalin-fixed, paraffin-embedded) staining of Human Liver tissue using Product # PA5-19471, anti-COX IV antibody. Primary antibody was used at a concentration of 1 µg/mL and exposed for 8 mins at room temp. The sample was pretreated using heat mediated antigen retrieval with Sodium Citrate Buffer (pH6/20mins). The detection method was a HRP conjugated polymer, DAB chromogen and the sample was counterstained with haematoxylin and mounted with DPX.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6 Metformin upregulates Tfam in C2C12 myoblasts but not when miR-378a-3p is inhibited. ( A - C ) Expression level of Tfam in C2C12 myoblasts. ( D ) Representative images of Western Blot showing TFAM and COXIV protein content. Mitochondria-related factors, such as ( E - G ) TFAM and ( H - J ) COXIV, were evaluated through Western Blot. All data is given as mean +- S.E.M. ( n = 3). * p = 0.014 versus Glc 25 mM, ** p = 0.002 versus Glc 25 mM.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 6 Hepatic mitochondrial protein content quantification by Western blot. Data are means +- SEM of 4 independent experiments. ATP5b, ATP synthase F1 subunit beta, mitochondrial; ATP6, ATP synthase F O subunit 6, mitochondrial; COX II, cytochrome c oxidase subunit II; COX IV, cytochrome c oxidase, subunit IV; ND5, NADH-ubiquinone oxidoreductase chain 5; TFAM, mitochondrial transcription factor A; sAC, soluble adenylyl cyclase; LC3b, microtubule-associated proteins 1A/1B light chain 3B. * indicates a statistically significant difference vs. Ctl; # indicates a statistically significant difference vs. I/R.