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Product Info Summary
| SKU: | A00002-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | IHC, WB, ELISA (Cap) |
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Product info
Product Name
Anti-TNF alpha Antibody Picoband®
SKU/Catalog Number
A00002-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-TNF alpha Antibody Picoband® catalog # A00002-3. Tested in ELISA, IHC, WB applications. This antibody reacts with Human. 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-TNF alpha Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00002-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human TNF alpha recombinant protein (Position: V77-L233).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00002-3 is reactive to TNF in Human
Observed Molecular Weight
17 kDa
Calculated molecular weight
25.6 kDa
Background of TNF
TNF (Tumor Necrosis Factor alpha) gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.
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
A00002-3 is guaranteed for IHC, WB, ELISA (Cap) 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
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
ELISA (Cap), 1-5μg/ml, -
Positive Control
WB: recombinant human TNF alpha protein
IHC: human spleen tissue, human tonsil tissue
Validation Images & Assay Conditions
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Western blot analysis of TNF alpha using anti-TNF alpha antibody (A00002-3).
Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours.
Lane 1: recombinant human TNF alpha protein 10 ng,
Lane 2: recombinant human TNF alpha protein 5 ng,
Lane 3: recombinant human TNF alpha protein 1 ng,
Lane 4: recombinant human TNF alpha protein 0.5 ng.
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-TNF alpha antigen affinity purified polyclonal antibody (A00002-3) 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 (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 TNF alpha at approximately 17 kDa.
Click image to see more details
IHC analysis of TNF alpha using anti-TNF alpha antibody (A00002-3).
TNF alpha was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-TNF alpha Antibody (A00002-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
IHC analysis of TNF alpha using anti-TNF alpha antibody (A00002-3).
TNF alpha was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-TNF alpha Antibody (A00002-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Specific Publications For Anti-TNF alpha Antibody Picoband® (A00002-3)
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Customer Reviews
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Customer Q&As
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18 Customer Q&As for Anti-TNF alpha Antibody Picoband®
Question
Is this A00002-3 anti-TNF alpha antibody reactive to the isotypes of TNF?
Verified Customer
Verified customer
Asked: 2020-03-12
Answer
The immunogen of A00002-3 anti-TNF alpha antibody is E. coli-derived human TNF alpha recombinant protein (Position: V77-L233). Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2020-03-12
Question
Our lab used your anti-TNF alpha antibody for WB on prostatic carcinoma in a previous experiment. I am using human, and We intend to use the antibody for ELISA next. I am interested in examining prostatic carcinoma as well as blood in our next experiment. Could you please give me some suggestion on which antibody would work the best for ELISA?
T. Parker
Verified customer
Asked: 2020-02-20
Answer
I looked at the website and datasheets of our anti-TNF alpha antibody and it seems that A00002-3 has been validated on human in both WB and ELISA. Thus A00002-3 should work for your application. Our Boster satisfaction guarantee will cover this product for ELISA in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for ELISA detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2020-02-20
Question
I see that the anti-TNF alpha antibody A00002-3 works with ELISA, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2020-02-03
Answer
You can find protocols for ELISA 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: 2020-02-03
Question
Dear Support; What is the positive control slides I can use for this A00002-3 Anti-TNF alpha Antibody in Immunohistochemistry paraffin embedded sections? Best regards.
Verified Customer
Verified customer
Asked: 2019-12-23
Answer
For product A00002-3 Anti-TNF alpha Antibody, we recommend lung tissues. For your further references kindly read Tissue Antigens 16:30-48 (1980). PubMed: 6162228.
Boster Scientific Support
Answered: 2019-12-23
Question
I have an inquiry; what is the source of immunizing protein for this A00002-3 Anti-TNF alpha Antibody?, was the mouse immunized with native protein or recombinant expressed in E. coli, or insect cells or mammalian cells? Thank you,
Verified Customer
Verified customer
Asked: 2019-12-04
Answer
We have the following information from the source : The A00002-3 Anti-TNF alpha Antibody, TNF-RII (p75/p80), clone MR2-1 were generated as described in the following article: Leeuwenberg, JFM et al; Slow release of soluble TNF-Receptors by monocytes in vitro. J Immunol 1994, 152: 4036. In this article you can see that sTNF-R75 was purified from culture supernatant of cells, transfected with the extracellular part of TNF-R75 by affinity chromatography on a TNF-coupled agarose column.
Boster Scientific Support
Answered: 2019-12-04
Question
Please see the WB image, lot number and protocol we used for blood using anti-TNF alpha antibody A00002-3. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2019-11-20
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: 2019-11-20
Question
I was wanting to use your anti-TNF alpha antibody for ELISA for human blood on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human blood identification?
Verified Customer
Verified customer
Asked: 2019-09-12
Answer
It shows on the product datasheet, A00002-3 anti-TNF alpha antibody has been tested for ELISA, WB on human tissues. We have an innovator award program that if you test this antibody and show it works in human blood in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-09-12
Question
Is a blocking peptide available for product anti-TNF alpha antibody (A00002-3)?
Verified Customer
Verified customer
Asked: 2019-09-06
Answer
We do provide the blocking peptide for product anti-TNF alpha antibody (A00002-3). If you would like to place an order for it please contact support@bosterbio.com and make a special request.
Boster Scientific Support
Answered: 2019-09-06
Question
Hello Boster: I want to test A00002-3 Anti-TNF alpha Antibody in mouse samples. You think it is ok? Please let me know. Many thanks
Verified Customer
Verified customer
Asked: 2018-12-05
Answer
This A00002-3 Anti-TNF alpha Antibody has been positively tested in Dog and Human species in our lab, but not tested in Mouse. Though, due to the high sequence homology between the protein
Boster Scientific Support
Answered: 2018-12-05
Question
We are currently using anti-TNF alpha antibody A00002-3 for human tissue, and we are happy with the WB results. The species of reactivity given in the datasheet says human. Is it likely that the antibody can work on pig tissues as well?
T. Edwards
Verified customer
Asked: 2018-11-28
Answer
The anti-TNF alpha antibody (A00002-3) has not been tested for cross reactivity specifically with pig 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 pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-11-28
Question
Can you help my question with product A00002-3, anti-TNF alpha antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2018-11-05
Answer
We do not recommend storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free A00002-3 anti-TNF alpha antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2018-11-05
Question
Will A00002-3 anti-TNF alpha antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
G. Collins
Verified customer
Asked: 2018-08-16
Answer
It shows on the product datasheet, A00002-3 anti-TNF alpha antibody as been validated on ELISA. 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: 2018-08-16
Question
My boss were satisfied with the WB result of your anti-TNF alpha antibody. However we have seen positive staining in leukocyte cell membrane using this antibody. Is that expected? Could you tell me where is TNF supposed to be expressed?
Verified Customer
Verified customer
Asked: 2017-11-30
Answer
Based on literature, leukocyte does express TNF. Generally TNF expresses in cell membrane. Regarding which tissues have TNF expression, here are a few articles citing expression in various tissues:
Blood, Pubmed ID: 15489334
Prostatic carcinoma, Pubmed ID: 8597870, 10205166
Boster Scientific Support
Answered: 2017-11-30
Question
Would anti-TNF alpha antibody A00002-3 work for ELISA with blood?
Verified Customer
Verified customer
Asked: 2017-08-15
Answer
According to the expression profile of blood, TNF is highly expressed in blood. So, it is likely that anti-TNF alpha antibody A00002-3 will work for ELISA with blood.
Boster Scientific Support
Answered: 2017-08-15
Question
you antibody to test anti-TNF alpha antibody A00002-3 on human blood for research purposes, then I may be interested in using anti-TNF alpha antibody A00002-3 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2017-06-22
Answer
The products we sell, including anti-TNF alpha antibody A00002-3, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2017-06-22
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for blood using anti-TNF alpha antibody A00002-3. Let me know if you need anything else.
J. Zhang
Verified customer
Asked: 2016-10-11
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2016-10-11
Question
We have observed staining in human blood. What should we do? Is anti-TNF alpha antibody supposed to stain blood positively?
Z. Collins
Verified customer
Asked: 2014-08-14
Answer
From what I have seen in literature blood does express TNF. From what I have seen in Uniprot.org, TNF is expressed in leukocyte, blood, prostatic carcinoma, among other tissues. Regarding which tissues have TNF expression, here are a few articles citing expression in various tissues:
Blood, Pubmed ID: 15489334
Prostatic carcinoma, Pubmed ID: 8597870, 10205166
Boster Scientific Support
Answered: 2014-08-14
Question
Do you have a BSA free version of anti-TNF alpha antibody A00002-3 available?
P. Wu
Verified customer
Asked: 2014-06-27
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-TNF alpha antibody A00002-3 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: 2014-06-27
