Anti-HAO1 Antibody Picoband™

Product Name

Anti-HAO1 Antibody Picoband™

View all Hydroxyacid Oxidase-1/HAO-1 Antibodies

SKU/Catalog Number

A09159-2

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-HAO1 Antibody Picoband™ catalog # A09159-2. Tested in ELISA, IHC, WB, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-HAO1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A09159-2)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

IgG

Immunogen

E.coli-derived human HAO1 recombinant protein (Position: K48-I370). Human HAO1 shares 90.4% and 90.7% amino acid (aa) sequence identity with mouse and rat HAO1, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

A09159-2 is reactive to Hao1 in Human, Mouse, Rat

Applications

A09159-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

41 kDa

Calculated molecular weight

52588 MW

Background of Hydroxyacid Oxidase-1/HAO-1

Hydroxyacid oxidase (glycolate oxidase) 1 is a protein that in humans is encoded by the HAO1 gene. This gene is one of three related genes that have 2-hydroxyacid oxidase activity yet differ in encoded protein amino acid sequence, tissue expression and substrate preference. Subcellular location of the encoded protein is the peroxisome. Specifically, this gene is expressed primarily in liver and pancreas and the encoded protein is most active on glycolate, a two-carbon substrate. The protein is also active on 2-hydroxy fatty acids. The transcript detected at high levels in pancreas may represent an alternatively spliced form or the use of a multiple near-consensus upstream polyadenylation site.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconstitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Mouse, Rat
Immunohistochemistry, 2-5 µg/ml, Mouse
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of HAO1 using anti-HAO1 antibody (A09159-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: mouse liver tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HAO1 antigen affinity purified polyclonal antibody (Catalog # A09159-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for HAO1 at approximately 41 kDa. The expected band size for HAO1 is at 41 kDa.

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Figure 2. IHC analysis of HAO1 using anti-HAO1 antibody (A09159-2).
HAO1 was detected in a paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HAO1 Antibody (A09159-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of HAO1 using anti-HAO1 antibody (A09159-2).
HAO1 was detected in a paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-HAO1 Antibody (A09159-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of SiHa cells using anti-HAO1 antibody (A09159-2).
Overlay histogram showing SiHa cells stained with A09159-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-HAO1 Antibody (A09159-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For Hao1 (Source: Uniprot.org, NCBI)

Alternative Names

(S)-2-hydroxy-acid oxidase; EC 1.1.3.15; Glycolate oxidase; GOX; GOX1MGC142227; GOXMGC142225; HAO1; HAO-1; HAOX1; hydroxyacid oxidase (glycolate oxidase) 1; hydroxyacid oxidase 1; Hydroxyacid Oxidase1; Hydroxyacid Oxidase-1 Hao1|GO, GOX, Gox1, Hao, Hao-1|hydroxyacid oxidase 1, liver|hydroxyacid oxidase 1|HAOX1|glycolate oxidase 1

For more info on Hao1, check out the Hao1 Infographic

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