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Product Info Summary
| SKU: | PA1848 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-B Raf/BRAF Antibody Picoband®
SKU/Catalog Number
PA1848
BA2958-2 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-B Raf/BRAF Antibody catalog # PA1848. Tested in Flow Cytometry， 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-B Raf/BRAF Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1848)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human B Raf, identical to the related rat and mouse sequences.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PA1848 is reactive to BRAF in Human, Mouse, Rat
Observed Molecular Weight
84 kDa
Calculated molecular weight
84.4 kDa
Background of BRAF
BRAF (v-raf murine sarcoma viral oncogene homolog B1) is a human gene that makes a protein called B-Raf, also known as ONCOGENE BRAF, BRAF1, RAFB1, Proto-oncogene B-Raf p94, v-Raf murine sarcoma viral oncogene homolog B1, proto-oncogene B-Raf, v-Raf murine sarcoma viral oncogene homolog B1, while the protein is more formally known as serine/threonine-protein kinase B-Raf. BRAF deduced 651-amino acid protein has a calculated molecular mass of 72.5 kD. Northern blot analysis detected transcripts of 10 and 13 kb in cerebrum, fetal brain, and placenta and transcripts of 2.6 and 4.5 kb in testis. Testis also showed lower expression of the 10- and 13-kb transcripts.B-Raf is a member of the Raf kinase family of serine/threonine-specific protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion.
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
PA1848 is guaranteed for Flow Cytometry, 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
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human MOLT4 whole cell, human RT4 whole cell, human K562 whole cell, rat brain tissue, rat PC-12 whole cell, mouse brain tissue
FCM: U937 cell
Validation Images & Assay Conditions
Click image to see more details
Western blot analysis of BRAF using anti-BRAF antibody (PA1848).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human MOLT4 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse brain tissue 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-BRAF antigen affinity purified polyclonal antibody (PA1848) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for BRAF at approximately 84 kDa. The expected band size for BRAF is at 84 kDa.
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Flow Cytometry analysis of U937 cells using anti-BRAF antibody (PA1848).
Overlay histogram showing U937 cells stained with PA1848 (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-BRAF Antibody (PA1848, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-B Raf/BRAF Antibody Picoband® (PA1848)
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Customer Q&As
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4 Customer Q&As for Anti-B Raf/BRAF Antibody Picoband®
Question
Is there a BSA free version of anti-B Raf/BRAF antibody PA1848 available?
Verified Customer
Verified customer
Asked: 2019-11-08
Answer
I appreciate your recent telephone inquiry. I can confirm that some lots of this anti-B Raf/BRAF antibody PA1848 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: 2019-11-08
Question
I see that the anti-B Raf/BRAF antibody PA1848 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2017-10-25
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2017-10-25
Question
I was wanting to use your anti-B Raf/BRAF antibody for WB for mouse colon carcinoma hepatoma on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for mouse colon carcinoma hepatoma identification?
G. Mitchell
Verified customer
Asked: 2017-08-01
Answer
You can see on the product datasheet, PA1848 anti-B Raf/BRAF antibody has been validated for WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse colon carcinoma hepatoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-08-01
Question
Will PA1848 anti-B Raf/BRAF antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
E. Jha
Verified customer
Asked: 2013-11-13
Answer
You can see on the product datasheet, PA1848 anti-B Raf/BRAF antibody as been tested 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: 2013-11-13
