Anti-Sema6A Antibody Picoband™

Product Name

Anti-Sema6A Antibody Picoband™

View all Semaphorin 6A Antibodies

SKU/Catalog Number

A06967-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Sema6A Antibody Picoband™ catalog # A06967-1. Tested in ELISA, Flow Cytometry, IHC, 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-Sema6A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06967-1)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Sema6A recombinant protein (Position: H31-N306).

*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

A06967-1 is reactive to Sema6a in Human, Mouse, Rat

Applications

A06967-1 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

114 kDa

Calculated molecular weight

114.433kDa

Background of Semaphorin 6A

SEMA6A belongs to a subfamily characterized by an extracellular semaphorin domain, a transmembrane domain, and a long cytoplasmic tail. Members of this class can repel sympathetic and dorsal root ganglion axons in vitro, consistent with a traditional role as guidance signals. However, the length of the cytoplasmic tail, which includes an EVL-binding site in SEMA6A and an Src-binding site in SEMA6B, suggests that these semaphorins may also function as receptors. SEMA6A is expressed in developing neural tissue and is required for proper development of the thalamocortical projection. SEMA6A directly links the Ena/VASP and the semaphorin protein families since the SEMA6A protein is capable of selective binding to the protein EVL (Ena/VASP-like protein).

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, 0.1-0.25μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Sema6A using anti-Sema6A antibody (A06967-1).
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 HEK293 whole cell lysates,
Lane 2: human HELA whole cell lysates,
Lane 3: human Raji whole cell lysates,
Lane 4: human U20S whole cell lysates,
Lane 5: human Jurkat whole cell lysates,
Lane 6: human CACO-2 whole cell lysates,
Lane 7: rat brain tissue lysates,
Lane 8: rat PC-12 whole cell lysates,
Lane 9: mouse brain tissue lysates,
Lane 10: mouse RAW264.7 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 rabbit anti-Sema6A antigen affinity purified polyclonal antibody (Catalog # A06967-1) 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-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 Sema6A at approximately 114KD. The expected band size for Sema6A is at 114KD.

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Figure 2. IHC analysis of Sema6A using anti-Sema6A antibody (A06967-1).
Sema6A was detected in paraffin-embedded section of human rectal 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-Sema6A Antibody (A06967-1) 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.

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Figure 3. Flow Cytometry analysis of HEPG2 cells using anti-Sema6A antibody (A06967-1).
Overlay histogram showing HEPG2 cells stained with A06967-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Sema6A Antibody (A06967-1, 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 (Red line) was also used as a control.

Gene/Protein Information For Sema6a (Source: Uniprot.org, NCBI)

Alternative Names

KIAA1368HT018; sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A; Sema VIA; SEMA; Sema6A; SEMA6A1; SEMA6A-1; Semaphorin 6A; semaphorin 6A-1; Semaphorin VIA; semaphorin-6A; semaphorin-6A-1; SEMAQ; VIA Sema6a|A730020P05Rik, AI851735, Sem, Sema6, Sema6A-1, Semaq, VIa, b2b997C, se|sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6A|semaphorin-6A|Mutant line 997|sema Q|sema VIA|semaphorin Q|semaphorin VIA|semaphorin-6A-1

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