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Product Info Summary
| SKU: | M02834-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-HMG4 Antibody Picoband® (monoclonal, 8H9)
View all HMGB3/HMG4 Antibodies
SKU/Catalog Number
M02834-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-HMG4 Antibody Picoband® (monoclonal, 8H9) catalog # M02834-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-HMG4 Antibody Picoband® (monoclonal, 8H9) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M02834-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
8H9
Isotype
Mouse IgG2b
Immunogen
A synthetic peptide corresponding to a sequence at the N-terminus of human HMG4, identical to the related mouse and rat sequences.
*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
M02834-1 is reactive to HMGB3 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
23 kDa
Calculated molecular weight
22.98kDa
Background of HMGB3/HMG4
High-mobility group protein B, also known as HMG4, is a protein that in humans is encoded by the HMGB3 gene. This gene encodes a member of a family of proteins containing one or more high mobility group DNA-binding motifs. The encoded protein plays an important role in maintaining stem cell populations, and may be aberrantly expressed in tumor cells. A mutation in this gene was associated with microphthalmia, syndromic 13. There are numerous pseudogenes of this gene on multiple chromosomes. Alternative splicing results in multiple transcript variants.
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.
Application & Images
Applications
M02834-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
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, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human Jurkat whole cell, human CACO-2 whole cell, human 293T whole cell, human THP-1 whole cell, human HepG2 whole cell, human MCF-7 whole cell, rat brain tissue, rat lung tissue, mouse kidney tissue, mouse brain tissue, mouse lung tissue, mouse ovary tissue
ICC/IF: Hela cell
FCM: Hela cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of HMGB3 using anti-HMGB3 antibody (M02834-1).
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 30ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human CACO-2 whole cell lysates,
Lane 4: human 293T whole cell lysates,
Lane 5: human THP-1 whole cell lysates,
Lane 6: human HepG2 whole cell lysates,
Lane 7: human MCF-7 whole cell 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 mouse anti-HMGB3 antigen affinity purified monoclonal antibody (Catalog # M02834-1) at 0.25 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 HMGB3 at approximately 23 kDa. The expected band size for HMGB3 is at 23 kDa.
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Figure 2. Western blot analysis of HMGB3 using anti-HMGB3 antibody (M02834-1).
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 30ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat lung tissue lysates,
Lane 3: mouse kidney tissue lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse lung tissue lysates,
Lane 6: mouse ovary 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 mouse anti-HMGB3 antigen affinity purified monoclonal antibody (Catalog # M02834-1) at 0.25 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 HMGB3 at approximately 23 kDa. The expected band size for HMGB3 is at 23 kDa.
Click image to see more details
Figure 3. IF analysis of HMGB3 using anti-HMGB3 antibody (M02834-1).
HMGB3 was detected in immunocytochemical section of Hela cell. 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 2μg/mL mouse anti-HMGB3 Antibody (M02834-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.
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Figure 4. Flow Cytometry analysis of Hela cells using anti-HMGB3 antibody (M02834-1).
Overlay histogram showing Hela cells stained with M02834-1 (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 mouse anti-HMGB3 Antibody (M02834-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 HMGB3 (Source: Uniprot.org, NCBI)
Gene Name
HMGB3
Full Name
High mobility group protein B3
Weight
22.98kDa
Superfamily
HMGB family
Alternative Names
"chromosomal protein, Nonhistone, HMG4 antibody|High mobility group (nonhistone chromosomal) protein 4 antibody|High mobility group box 3 antibody|High mobility group protein 2a antibody|High mobility group protein 4 antibody|High mobility group protein B3 antibody| High mobility group protein HMG4 antibody|HMG 4 antibody|HMG-2a antibody|HMG-4 antibody|HMG2A antibody|HMGB 3 antibody|HMGB3 antibody| HMGB3_HUMAN antibody|MGC90319 antibody|Non histone chromosomal protein antibody|Nonhistone chromosomal protein HMG4 antibody" HMGB3 HMG-2a, HMG-4, HMG2A, HMG4 high mobility group box 3 high mobility group protein B3|high mobility group protein 2a|high-mobility group (nonhistone chromosomal) protein 4
*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 HMGB3, check out the HMGB3 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 HMGB3: 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-HMG4 Antibody Picoband® (monoclonal, 8H9) (M02834-1)
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Customer Q&As
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1 Customer Q&As for Anti-HMG4 Antibody Picoband® (monoclonal, 8H9)
Question
I would like using your anti-HMG4 antibody (monoclonal, 8H9) for negative regulation of b cell differentiation studies. Has this antibody been tested with western blotting on u20s cells? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-12-04
Answer
We appreciate your inquiry. This M02834-1 anti-HMG4 antibody (monoclonal, 8H9) is validated on human placenta tissue, hepg2 whole cell lysates, k562 whole cell lysates, mouse liver tissue, rat heart tissue, kidney tissue, u20s cells, siha cells. It is guaranteed to work for Flow Cytometry, IF, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-12-04
