Product Info Summary
SKU: | A06559-1 |
---|---|
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-Semaphorin 3B/SEMA3B Antibody Picoband™
View all Semaphorin 3B Antibodies
SKU/Catalog Number
A06559-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Semaphorin 3B/SEMA3B Antibody Picoband™ catalog # A06559-1. 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-Semaphorin 3B/SEMA3B Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06559-1)
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 Semaphorin 3B/SEMA3B recombinant protein (Position: Y55-R624).
*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
A06559-1 is reactive to SEMA3B in Human, Mouse, Rat
Applications
A06559-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
83 kDa
Calculated molecular weight
104553 MW
Background of Semaphorin 3B
Semaphorin-3B is a protein that in humans is encoded by the SEMA3B gene. It is mapped to 3p21.31. The semaphorin/collapsin family of molecules plays a critical role in the guidance of growth cones during neuronal development. The secreted protein encoded by this gene family member is important in axonal guidance and has been shown to act as a tumor suppressor by inducing apoptosis.
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, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Direct ELISA, 0.1-0.5μg/ml
Validation Images & Assay Conditions
![A06559 1 SEMA3B primary antibodies WB testing 1 A06559 1 SEMA3B primary antibodies WB testing 1](https://www.bosterbio.com/media/catalog/product/A/0/A06559-1-SEMA3B-primary-antibodies-WB-testing-1.jpg)
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Figure 1. Western blot analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
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 placenta tissue lysates
Lane 2: human U2OS whole cell lysates
Lane 3: human A549 whole cell lysates
Lane 4: human U-87MG 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-SEMA3B antigen affinity purified polyclonal antibody (Catalog # A06559-1) 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 SEMA3B at approximately 83KD. The expected band size for SEMA3B is at 83KD.
![A06559 1 SEMA3B primary antibodies WB testing 2 A06559 1 SEMA3B primary antibodies WB testing 2](https://www.bosterbio.com/media/catalog/product/A/0/A06559-1-SEMA3B-primary-antibodies-WB-testing-2.jpg)
Click image to see more details
Figure 2. Western blot analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
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: rat thymus tissues lysates
Lane 2: rat kidney tissue lysates
Lane 3: mouse thymus tissue lysates
Lane 4: mouse lung tissue lysates
Lane 5: mouse small intestine tissue lysates
Lane 6: mouse kidney tissue lysates
Lane 7: mouse HEPA1-6 whole cel 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-SEMA3B antigen affinity purified polyclonal antibody (Catalog # A06559-1) 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 SEMA3B at approximately 83KD. The expected band size for SEMA3B is at 83KD.
![a06559 1 sema3b primary antibodies ihc testing 3 a06559 1 sema3b primary antibodies ihc testing 3](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-3.jpg)
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Figure 3. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies ihc testing 4 a06559 1 sema3b primary antibodies ihc testing 4](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-4.jpg)
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Figure 4. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in paraffin-embedded section of human colon 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies ihc testing 5 a06559 1 sema3b primary antibodies ihc testing 5](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-5.jpg)
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Figure 5. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in paraffin-embedded section of mouse brain 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies ihc testing 6 a06559 1 sema3b primary antibodies ihc testing 6](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-6.jpg)
Click image to see more details
Figure 6. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in paraffin-embedded section of mouse brain 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies ihc testing 7 a06559 1 sema3b primary antibodies ihc testing 7](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-7.jpg)
Click image to see more details
Figure 7. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in paraffin-embedded section of rat brain 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies ihc testing 8 a06559 1 sema3b primary antibodies ihc testing 8](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-ihc-testing-8.jpg)
Click image to see more details
Figure 8. IHC analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in paraffin-embedded section of rat brain 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies if testing 9 a06559 1 sema3b primary antibodies if testing 9](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-if-testing-9.jpg)
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Figure 9. IF analysis of SEMA3B using anti-SEMA3B antibody (A06559-1).
SEMA3B was detected in immunocytochemical section of A431 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-SEMA3B Antibody (A06559-1) 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.
![a06559 1 sema3b primary antibodies fcm testing 10 a06559 1 sema3b primary antibodies fcm testing 10](https://www.bosterbio.com/media/catalog/product/a/0/a06559-1-sema3b-primary-antibodies-fcm-testing-10.jpg)
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Figure 10. Flow Cytometry analysis of A549 cells using anti-SEMA3B antibody (A06559-1).
Overlay histogram showing A549 cells stained with A06559-1 (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-SEMA3B Antibody (A06559-1,1μg/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 SEMA3B (Source: Uniprot.org, NCBI)
Gene Name
SEMA3B
Full Name
Semaphorin-3B
Weight
104553 MW
Superfamily
semaphorin family
Alternative Names
(semaphorin) 3B; LUCA-1; Sema A(V); sema domain, immunoglobulin domain (Ig), short basic domain, secreted; Sema V; SemA; SEMA3B; SEMA5; SEMA5FLJ34863; SEMAA; Semaphorin 3B; semaphorin A; semaphorin V; semaphorin-3B; semaphorin-V; SEMAV SEMA3B LUCA-1, SEMA5, SEMAA, SemA, semaV semaphorin 3B semaphorin-3B|sema A(V)|sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3B|semaphorin A|semaphorin-V
*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 SEMA3B, check out the SEMA3B Infographic
![SEMA3B infographic](/media/images/gene-infographic-example.jpg)
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for SEMA3B: 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-Semaphorin 3B/SEMA3B Antibody Picoband™ (A06559-1)
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Customer Q&As
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4 Customer Q&As for Anti-Semaphorin 3B/SEMA3B Antibody Picoband™
Question
See below the WB image, lot number and protocol we used for brain muscle using anti-Semaphorin 3B/SEMA3B antibody A06559-1. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2020-03-17
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2020-03-17
Question
Will A06559-1 anti-Semaphorin 3B/SEMA3B antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-02-14
Answer
You can see on the product datasheet, A06559-1 anti-Semaphorin 3B/SEMA3B antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2019-02-14
Question
We are currently using anti-Semaphorin 3B/SEMA3B antibody A06559-1 for human tissue, and we are happy with the ELISA results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on dog tissues as well?
Verified Customer
Verified customer
Asked: 2018-02-28
Answer
The anti-Semaphorin 3B/SEMA3B antibody (A06559-1) has not been tested for cross reactivity specifically with dog tissues, but 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-02-28
Question
Do you have a BSA free version of anti-Semaphorin 3B/SEMA3B antibody A06559-1 available?
Verified Customer
Verified customer
Asked: 2017-08-17
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Semaphorin 3B/SEMA3B antibody A06559-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2017-08-17