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Product Info Summary
| SKU: | PA1079 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-TNF alpha Antibody Picoband®
SKU/Catalog Number
PA1079
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-TNF alpha Antibody catalog # PA1079. Tested in Flow Cytometry, IHC, 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-TNF alpha Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1079)
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 at the C-terminus of human TNF alpha, different from the related mouse sequence by five amino acids, and rat sequence by seven amino acids.
*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
PA1079 is reactive to TNF in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
25 kDa
Calculated molecular weight
25503 MW
Background of TNF-alpha
TNF alpha (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
PA1079 is guaranteed for Flow Cytometry, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, By Heat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human U937 whole cell, rat spleen tissue, rat C6 whole cell, mouse spleen tissue
IHC: human B lymphocytic tumor tissue
FCM: CACO-2 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of TNF alpha using anti-TNF alpha antibody (PA1079).
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 U937 whole cell lysates,
Lane 2: rat spleen tissue lysates,
Lane 3: rat C6 whole cell lysates,
Lane 4: mouse spleen 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-TNF alpha antigen affinity purified polyclonal antibody (Catalog # PA1079) 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 TNF alpha at approximately 25 kDa. The expected band size for TNF alpha is at 26 kDa.
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Figure 2. IHC analysis of TNF alpha using anti-TNF alpha antibody (PA1079).
TNF alpha was detected in a paraffin-embedded section of human B lymphocytic tumor 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 (PA1079) 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.
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Figure 3. Flow Cytometry analysis of CACO-2 cells using anti-TNF alpha antibody (PA1079).
Overlay histogram showing CACO-2 cells stained with PA1079 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF alpha Antibody (PA1079, 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 (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For TNF (Source: Uniprot.org, NCBI)
Gene Name
TNF
Full Name
Tumor necrosis factor
Weight
25503 MW
Superfamily
Tumor necrosis factor family
Alternative Names
Tumor necrosis factor; Cachectin; TNF-alpha; Tumor necrosis factor ligand superfamily member 2; TNF-a; Tumor necrosis factor, membrane form; N-terminal fragment; NTF; Intracellular domain 1; ICD1; Intracellular domain 2; ICD2; C-domain 1; C-domain 2; Tumor necrosis factor, soluble form; TNF; TNFA; TNFSF2 TNF DIF-alpha, TNFA, TNFSF2, TNLG1F, TNF tumor necrosis factor tumor necrosis factor|APC1 protein|TNF, macrophage-derived|TNF, monocyte-derived|TNF-a|tumor necrosis factor ligand 1F|tumor necrosis factor ligand superfamily member 2|tumor necrosis factor-alpha
*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 TNF, check out the TNF 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 TNF: 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-TNF alpha Antibody Picoband® (PA1079)
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Customer Q&As
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16 Customer Q&As for Anti-TNF alpha Antibody Picoband®
Question
We are currently using anti-TNF alpha antibody PA1079 for rat tissue, and we are happy with the IF results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on bovine tissues as well?
Verified Customer
Verified customer
Asked: 2020-05-01
Answer
The anti-TNF alpha antibody (PA1079) has not been validated for cross reactivity specifically with bovine tissues, though 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-05-01
Question
I see that the anti-TNF alpha antibody PA1079 works with IF, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2020-04-17
Answer
You can find protocols for IF on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2020-04-17
Question
Do you have a BSA free version of anti-TNF alpha antibody PA1079 available?
Verified Customer
Verified customer
Asked: 2020-02-25
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-TNF alpha antibody PA1079 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: 2020-02-25
Question
My lab would like using your anti-TNF alpha antibody for positive regulation of membrane protein ectodomain proteolysis studies. Has this antibody been tested with western blotting on a431 whole cell lysates? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-12-23
Answer
We appreciate your inquiry. This PA1079 anti-TNF alpha antibody is tested on human placenta tissue, hela whole cell lysates, a431 whole cell lysates, a549 whole cell lysates, k562 whole cell lysates. It is guaranteed to work for IF, IHC-P, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-12-23
Question
Our lab want to know about to test anti-TNF alpha antibody PA1079 on mouse prostatic carcinoma for research purposes, then I may be interested in using anti-TNF alpha antibody PA1079 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
W. Wu
Verified customer
Asked: 2019-11-18
Answer
The products we sell, including anti-TNF alpha antibody PA1079, 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: 2019-11-18
Question
Is a blocking peptide available for product anti-TNF alpha antibody (PA1079)?
Verified Customer
Verified customer
Asked: 2019-06-13
Answer
We do provide the blocking peptide for product anti-TNF alpha antibody (PA1079). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-06-13
Question
See attached the WB image, lot number and protocol we used for prostatic carcinoma using anti-TNF alpha antibody PA1079. Please let me know if you require anything else.
R. Thomas
Verified customer
Asked: 2019-06-12
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: 2019-06-12
Question
We ordered your anti-TNF alpha antibody for IHC-P on leukocyte a few years ago. I am using rat, and We are going to use the antibody for WB next. I am interested in examining leukocyte as well as prostatic carcinoma in our next experiment. Could you please give me some suggestion on which antibody would work the best for WB?
J. Krishna
Verified customer
Asked: 2019-05-31
Answer
I viewed the website and datasheets of our anti-TNF alpha antibody and it appears that PA1079 has been tested on rat in both IHC-P and WB. Thus PA1079 should work for your application. Our Boster satisfaction guarantee will cover this product for WB in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for WB detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-05-31
Question
My question regarding product PA1079, 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: 2018-08-06
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 PA1079 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: 2018-08-06
Question
Our lab were well pleased with the WB result of your anti-TNF alpha antibody. However we have observed positive staining in blood cell membrane using this antibody. Is that expected? Could you tell me where is TNF supposed to be expressed?
Verified Customer
Verified customer
Asked: 2017-07-24
Answer
From literature, blood 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: 2017-07-24
Question
We have observed staining in human leukocyte. Are there any suggestions? Is anti-TNF alpha antibody supposed to stain leukocyte positively?
Verified Customer
Verified customer
Asked: 2017-07-19
Answer
According to literature leukocyte does express TNF. According to 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: 2017-07-19
Question
I was wanting to use your anti-TNF alpha antibody for IF for mouse prostatic carcinoma 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 mouse prostatic carcinoma identification?
L. Thomas
Verified customer
Asked: 2016-12-01
Answer
It shows on the product datasheet, PA1079 anti-TNF alpha antibody has been validated for IF, IHC-P, ICC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse prostatic carcinoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2016-12-01
Question
Is this PA1079 anti-TNF alpha antibody reactive to the isotypes of TNF?
M. Edwards
Verified customer
Asked: 2015-06-18
Answer
The immunogen of PA1079 anti-TNF alpha antibody is A synthetic peptide corresponding to a sequence at the C-terminus of human TNF(201-233aa QLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL), different from the related mouse sequence by five amino acids, and rat sequence by seven amino acids. 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: 2015-06-18
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for prostatic carcinoma using anti-TNF alpha antibody PA1079. Let me know if you need anything else.
A. Collins
Verified customer
Asked: 2015-02-18
Answer
We appreciate 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: 2015-02-18
Question
Does anti-TNF alpha antibody PA1079 work for IF with prostatic carcinoma?
E. Anderson
Verified customer
Asked: 2014-03-05
Answer
According to the expression profile of prostatic carcinoma, TNF is highly expressed in prostatic carcinoma. So, it is likely that anti-TNF alpha antibody PA1079 will work for IF with prostatic carcinoma.
Boster Scientific Support
Answered: 2014-03-05
Question
Would PA1079 anti-TNF alpha antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
K. Collins
Verified customer
Asked: 2013-07-12
Answer
It shows on the product datasheet, PA1079 anti-TNF alpha antibody as been validated on IF. 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: 2013-07-12
