My Western blot shows two prominent bands for ACOX2. Is the antibody non-specific?

No, this is the expected result and a sign of high specificity. ACOX2 is synthesized as a precursor protein that is cleaved inside the peroxisome into two active subunits of approximately 50 kDa and 20 kDa. Detecting these two bands indicates that the antibody recognizes the mature, functional form of the enzyme.

I am performing IHC on liver tissue and see some background. Do you have any suggestions for improving the signal-to-noise ratio?

High background in IHC can often be reduced with a robust blocking step. We recommend using a blocking buffer containing 5% normal serum from the same species as your secondary antibody (e.g., normal goat serum if using a goat anti-mouse secondary). You can also try titrating the primary antibody to a higher dilution to find the optimal concentration that maximizes specific signal while minimizing background.

Mouse Anti-ACOX2 Recombinant Antibody (V2-179361) (CBMAB-A0662-YC)

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Datasheet

SDS

Request for COA

Datasheet Target References Q & As Review & reward Protocols Associated Products

Basic Information

Host Animal

Mouse

Clone

V2-179361

Application

ELISA, WB

Immunogen

ACOX2 (NP_003491, 582 a.a. ~ 681 a.a) partial recombinant protein with GST tag.

Specificity

Human

Antibody Isotype

IgG1, κ

Clonality

Monoclonal

Application Notes

The COA includes recommended starting dilutions, optimal dilutions should be determined by the end user.

Formulations & Storage [For reference only, actual COA shall prevail!]

Format

PBS, pH 7.2

Buffer

PBS, pH7.4

Preservative

None

Concentration

Batch dependent

Storage

Store at 4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

More Infomation

Target

Full Name

acyl-Coenzyme A oxidase 2, branched chain

Introduction

ACOX2 belongs to the acyl-CoA oxidase family. It encodes the branched-chain acyl-CoA oxidase which is involved in the degradation of long branched fatty acids and bile acid intermediates in peroxisomes. Deficiency of this enzyme results in the accumulatio

Entrez Gene ID

8309

UniProt ID

Q99424

Alternative Names

Acyl-CoA Oxidase 2; 3-Alpha,7-Alpha,12-Alpha-Trihydroxy-5-Beta-Cholestanoyl-CoA 24-Hydroxylase; Trihydroxycoprostanoyl-CoA Oxidase; 3-Alpha,7-Alpha,12-Alpha-Trihydroxy-5-Beta-Cholestanoyl-CoA Oxidase; Acyl-Coenzyme A Oxidase 2, Branched Chain; Acyl-CoA Ox

Function

Oxidizes the CoA esters of the bile acid intermediates di- and tri-hydroxycholestanoic acids. Capable of oxidizing short as well as long chain 2-methyl branched fatty acids (By similarity).

Biological Process

Bile acid biosynthetic process
Fatty acid beta-oxidation using acyl-CoA oxidase
Lipid homeostasis
Positive regulation of cell death
Positive regulation of response to oxidative stress
Protein localization
Very long-chain fatty acid metabolic process

Cellular Location

Peroxisome

Involvement in disease

Congenital bile acid synthesis defect 6 (CBAS6): An inborn error of bile acid synthesis characterized by abnormally increased liver enzymes, hypolipidemia and low cholesterol, vitamin D deficiency, elevated plasma and urinary levels of C27 intermediate bile acids 3alpha,7alpha-dihydroxy-5beta-cholestanoic acid (DHCA) and 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestanoic acid (THCA). Serum levels of phytanic and pristanic acids are normal. Clinical features include liver fibrosis, mild ataxia, delayed development, and cognitive impairment. Liver histology shows many thin fibrous septa, swollen hepatocytes, glycogenated nuclei, and focal acinar transformation, consistent with hepatocellular injury and regeneration, without signs of obvious cholestasis, cholate stasis, or steatosis. CBAS6 transmission pattern is consistent with autosomal recessive inheritance.

Okamura, M., Ueno, T., Tanaka, S., Murata, Y., Kobayashi, H., Miyamoto, A., ... & Fukuda, N. (2021). Increased expression of acyl-CoA oxidase 2 in the kidney with plasma phytanic acid and altered gut microbiota in spontaneously hypertensive rats. Hypertension Research, 44(6), 651-661.

Zhang, Q., Zhang, Y., Sun, S., Wang, K., Qian, J., Cui, Z., ... & Zhou, J. (2021). ACOX2 is a prognostic marker and impedes the progression of hepatocellular carcinoma via PPARα pathway. Cell Death & Disease, 12(1), 1-12.

Alonso-Pena, M., HERMANS, H., Herraez, E., Briz, O., MONTE, M., Sanchez-Martin, M., ... & MARIN, J. (2019, April). Genetic and pathophysiological factors leading to deficient acyl-CoA oxidase 2 (ACOX2) activity in hepatocytes, an alteration which causes oxidative and endoplasmic reticulum stress in liver cells. In Journal of Hepatology (Vol. 70, pp. E579-E579). PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS: ELSEVIER SCIENCE BV.

Ferdinandusse, S., Denis, S., van Roermund, C. W., Preece, M. A., Koster, J., Ebberink, M. S., ... & Wanders, R. J. (2018). A novel case of ACOX2 deficiency leads to recognition of a third human peroxisomal acyl-CoA oxidase. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease, 1864(3), 952-958.

Zhou, X., & Wang, H. (2017). ACOX2 deficiency in primary malignant cardiac tumors. Proceedings of the National Academy of Sciences, 114(18), E3590-E3591.

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Please try the standard protocols which include: protocols, troubleshooting and guide.

Enzyme-linked Immunosorbent Assay (ELISA)
Flow Cytometry
Immunofluorescence (IF)
Immunohistochemistry (IHC)
Immunoprecipitation (IP)
Western Blot (WB)
Enzyme-Linked Immunospot (ELISpot)
Proteogenomics
Other Protocols

Related Products

Mouse Anti-ACOX2 Recombinant Antibody (V2-179363) (CAT#: CBMAB-A0664-YC)

Mouse Anti-ACOX2 Recombinant Antibody (V2-179362) (CAT#: CBMAB-A0663-YC)

Isotype control

For research use only. Not intended for any clinical use.

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We also offer labeled antibodies developed using our catalog antibody products and nonfluorescent conjugates (HRP, AP, Biotin, etc.) or fluorescent conjugates (Alexa Fluor, FITC, TRITC, Rhodamine, Texas Red, R-PE, APC, Qdot Probes, Pacific Dyes, etc.).

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