Anti-B7H4/VTCN1 Antibody Picoband™

Product Name

Anti-B7H4/VTCN1 Antibody Picoband™

View all B7-H4 Antibodies

SKU/Catalog Number

A02821-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-B7H4/VTCN1 Antibody Picoband™ catalog # A02821-3. 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-B7H4/VTCN1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02821-3)

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 B7H4/VTCN1 recombinant protein (Position: T57-A258).

*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

A02821-3 is reactive to VTCN1 in Human, Mouse, Rat

Applications

A02821-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

31 kDa

Calculated molecular weight

30.878kDa

Background of B7-H4

V-set domain-containing T-cell activation inhibitor 1 is a protein that in humans is encoded by the VTCN1 gene. It is mapped to 1p13.1-p12. This gene encodes a protein belonging to the B7 costimulatory protein family. Proteins in this family are present on the surface of antigen-presenting cells and interact with ligand bound to receptors on the surface of T cells. Studies have shown that high levels of the encoded protein has been correlated with tumor progression. A pseudogene of this gene is located on chromosome 20. Multiple transcript variants encoding different isoforms have been found for this gene.

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

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, 0.25-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry, 1-3μg/1x10⁶ cells
Direct ELISA, 0.1-0.5μg/ml

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of VTCN1 using anti-VTCN1 antibody (A02821-3).
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 MCF-7 whole cell lysates,
Lane 2: human Hacat whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: rat liver tissue lysates,
Lane 5: rat RH35 whole cell lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: moue liver tissue lysates,
Lane 8: mouse HEPA1-6 whole cell 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-VTCN1 antigen affinity purified polyclonal antibody (Catalog # A02821-3) 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 VTCN1 at approximately 75 kDa. The expected band size for VTCN1 is at 31 kDa.

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Figure 2. IHC analysis of VTCN1 using anti-VTCN1 antibody (A02821-3).
VTCN1 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-VTCN1 Antibody (A02821-3) 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.

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Figure 3. IF analysis of VTCN1 using anti-VTCN1 antibody (A02821-3).
VTCN1 was detected in immunocytochemical section of U20S 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 2μg/mL rabbit anti-VTCN1 Antibody (A02821-3) 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.

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Figure 4. Flow Cytometry analysis of U20S cells using anti-VTCN1 antibody (A02821-3).
Overlay histogram showing U20S cells stained with A02821-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-VTCN1 Antibody (A02821-3,1μg/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 (Red line) was also used as a control.

Gene/Protein Information For VTCN1 (Source: Uniprot.org, NCBI)

Alternative Names

B7h.5; B7H4; B7-H4; B7H4T-cell costimulatory molecule B7x; B7S1; B7S1VCTN1; B7x; B7XPRO1291; FLJ22418; Immune costimulatory protein B7-H4; Protein B7S1; T cell costimulatory molecule B7x; V-set domain containing T cell activation inhibitor 1; V-set domain-containing T-cell activation inhibitor 1; Vtcn1 VTCN1 B7-H4, B7H4, B7S1, B7X, B7h.5, PRO1291, VCTN1 V-set domain containing T cell activation inhibitor 1 V-set domain-containing T-cell activation inhibitor 1|B7 family member, H4|B7 homolog 4|B7 superfamily member 1|T cell costimulatory molecule B7x|immune costimulatory protein B7-H4

For more info on VTCN1, check out the VTCN1 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 VTCN1: 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].

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