Product Info Summary
SKU: | A01909-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-HGD Antibody Picoband™
SKU/Catalog Number
A01909-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-HGD Antibody Picoband™ catalog # A01909-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-HGD Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01909-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
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
A01909-1 is reactive to HGD in Human, Mouse, Rat
Applications
A01909-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
50 kDa
Calculated molecular weight
40895 MW
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.
Assay dilution & Images
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/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,0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section),0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 5μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Direct ELISA,0.1-0.5μg/ml
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of HGD using anti-HGD antibody (A01909-1).
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 50ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse kidney tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-HGD antigen affinity purified polyclonal antibody (Catalog # A01909-1) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for HGD at approximately 50KD. The expected band size for HGD is at 50KD.
Click image to see more details
Figure 2. IHC analysis of HGD using anti-HGD antibody (A01909-1).
HGD was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-HGD Antibody (A01909-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of HGD using anti-HGD antibody (A01909-1).
HGD was detected in paraffin-embedded section of human prostatic cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-HGD Antibody (A01909-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IF analysis of HGD using anti-HGD antibody (A01909-1).
HGD was detected in immunocytochemical section of CACO-2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-HGD Antibody (A01909-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 5. Flow Cytometry analysis of HEPG2 cells using anti-HGD antibody (A01909-1).
Overlay histogram showing HEPG2 cells stained with A01909-1 (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-HGD Antibody (A01909-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For HGD (Source: Uniprot.org, NCBI)
Gene Name
HGD
Full Name
Homogentisate 1,2-dioxygenase
Weight
40895 MW
Superfamily
homogentisate dioxygenase family
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
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on HGD, check out the HGD Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for HGD: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-HGD Antibody Picoband™ (A01909-1)
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Customer Q&As
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1 Customer Q&As for Anti-HGD Antibody Picoband™
Question
We are currently using anti-HGD antibody A01909-1 for human tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on feline tissues as well?
Verified Customer
Verified customer
Asked: 2017-05-26
Answer
The anti-HGD antibody (A01909-1) has not been validated for cross reactivity specifically with feline tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-05-26