Anti-HGD Antibody Picoband™ (monoclonal, 2F11E1)

Product Name

Anti-HGD Antibody Picoband™ (monoclonal, 2F11E1)

View all HGD Antibodies

SKU/Catalog Number

M01909-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-HGD Antibody Picoband™ (monoclonal, 2F11E1) catalog # M01909-1. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human.

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-HGD Antibody Picoband™ (monoclonal, 2F11E1) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01909-1)

Host

Mouse

Contents

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

Clonality

Monoclonal

Clone Number

2F11E1

Isotype

Mouse IgG2b

Immunogen

E. coli-derived human HGD recombinant protein (Position: D374-N445).

*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

M01909-1 is reactive to HGD in Human

Applications

M01909-1 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

50 kDa

Calculated molecular weight

49.964kDa

Background of HGD

The HGD gene encodes homogentisate 1,2-dioxygenase (HGD), an enzyme involved in the catabolism of phenylalanine and tyrosine. This enzyme is involved in the catabolism of the amino acids tyrosine and phenylalanine. Mutations in this gene are the cause of the autosomal recessive metabolism disorder alkaptonuria. This gene is mapped to chromosome 3q21-q23 by a preliminary PCR screen of hamster/human somatic cell hybrid genomic DNA samples and by fluorescence in situ hybridization.

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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Reconsitution

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, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of HGD using anti-HGD antibody (M01909-1).
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: human hepatocellular carcinoma paracancerous tissue (HCCP) 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 mouse anti-HGD antigen affinity purified monoclonal antibody (Catalog # M01909-1) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for HGD at approximately 50 kDa. The expected band size for HGD is at 50 kDa.

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Figure 2. IHC analysis of HGD using anti-HGD antibody (M01909-1).
HGD was detected in a paraffin-embedded section of human liver cancer 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 mouse anti-HGD Antibody (M01909-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of CACO-2 cells using anti-HGD antibody (M01909-1).
Overlay histogram showing CACO-2 cells stained with M01909-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HGD Antibody (M01909-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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

Alternative Names

AKU; EC 1.13.11.5; HGOFLJ94126; homogentisate 1,2-dioxygenase (homogentisate oxidase); homogentisate 1,2-dioxygenase; homogentisate oxidase; Homogentisate oxygenase; Homogentisic acid oxidase; homogentisicase HGD AKU, HGO homogentisate 1,2-dioxygenase homogentisate 1,2-dioxygenase|homogentisate oxidase|homogentisate oxygenase|homogentisic acid oxidase|homogentisicase

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