Anti-TNF alpha 抗体 [52B83] (ab1793)

Reviews (13)Q&A (33)References (101)

製品の概要

  • 製品名

    Anti-TNF alpha antibody [52B83]
    TNF alpha 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [52B83] to TNF alpha

  • 由来種

    Mouse

  • 特異性

    This antibody detects both natural and recombinant TNFa. It does not cross-react with TNF beta or lymphotoxin. It reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. It does not react with receptor-bound TNF-alpha. Non-specific background staining is observed in connective tissues.

  • アプリケーション

    適用あり: Flow Cyt, IHC-Fr, ICC/IF, ELISA, WB, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Guinea pig, Human, Chimpanzee, Zebrafish, Cynomolgus monkey, Rhesus monkey, Apteronotus leptorhynchus

  • 免疫原

    Full length native protein (purified) (Human).

  • ポジティブ･コントロール

  • 特記事項

    Dilution of 20-200 times is useful for IHC on paraffin embedded tissues and frozen sections; flow cytometry and Western blotting. Useful for staining classic paraffin embedded tissues without antigen retrieval.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab1793 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt 1/10.

(methanol fixed cells)

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/50.

Fixed in acetone for 10 minutes
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 17 kDa.

TNF-alpha is normally secreted as a homotrimer with a molecular mass of 52 kDa. Monomeric TNF-alpha  is 17.4 kDa

A reduced sample treatment and 15% SDS-Page was used.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能

    Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation.

  • 関連疾患

    Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis).

  • 配列類似性

    Belongs to the tumor necrosis factor family.

  • 翻訳後修飾

    The soluble form derives from the membrane form by proteolytic processing.
    The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
    O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid.

  • 細胞内局在

    Secreted and Cell membrane.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • APC1 antibody
    • APC1 protein antibody
    • Cachectin antibody
    • DIF antibody
    • Differentiation inducing factor antibody
    • Macrophage cytotoxic factor antibody
    • Tnf antibody
    • TNF superfamily member 2 antibody
    • TNF superfamily, member 2 antibody
    • TNF, macrophage derived antibody
    • TNF, monocyte derived antibody
    • TNF-a antibody
    • TNF-alpha antibody
    • TNFA antibody
    • TNFA_HUMAN antibody
    • TNFSF2 antibody
    • Tumor necrosis factor (TNF superfamily member 2) antibody
    • Tumor necrosis factor alpha antibody
    • Tumor necrosis factor antibody
    • Tumor necrosis factor ligand superfamily member 2 antibody
    • Tumor Necrosis Factor, Membrane Form antibody
    • Tumor necrosis factor, soluble form antibody
    see all

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNF alpha antibody [52B83] (ab1793)This image is courtesy of an Abreview submitted by Dr Robert Domitrovic
    ab1793 staining TNF alpha in mouse liver tissue sections, from mice intoxicated with carbon tetrachloride for 7 weeks, by IHC-P (Formaldehyde-fixed, Paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 1% BSA for 30 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/50 in 1% BSA) at 4°C for 12 hours. An undiluted HRP-conjugated secondary antibody was used.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    ab1793 staining TNFα in RAW 264.7 cells treated with (R,S)-rolipram (ab120029), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R,S)-rolipram, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120029 ((R,S)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Flow Cytometry - Anti-TNF alpha antibody [52B83] (ab1793)
    Flow Cytometry - Anti-TNF alpha antibody [52B83] (ab1793)
    Overlay histogram showing RAW 264.7 cells stained with ab1793 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1793, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in RAW 264.7 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Immunohistochemistry (Frozen sections) - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunohistochemistry (Frozen sections) - Anti-TNF alpha antibody [52B83] (ab1793)This image is courtesy of an anonymous Abreview
    ab1793 staining TNF alpha in Mouse duodenum and pancreatic cancer associated stroma sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were stained by DAPI.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNF alpha antibody [52B83] (ab1793)This image is courtesy of an anonymous Abreview
    ab1793 staining TNF alpha in Mouse pancreatic neoplasia tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with a MOM kit for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An undiluted Biotin-conjugated Goat anti-mouse IgG polyclonal (MOM kit) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    ab1793 staining TNFα in RAW 264.7 cells treated with (R)-(-)-rolipram (ab120031), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R)-(-)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120031 ((R)-(-)--rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody [52B83] (ab1793)

    ab1793 staining TNFα in RAW 264.7 cells treated with (S)-(+)-rolipram (ab120030), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (S)-(+)-rolipram , as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120030 ((S)-(+)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab1793(5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

参考文献

This product has been referenced in:

  • Jiang R  et al. Kaempferol protects chondrogenic ATDC5 cells against inflammatory injury triggered by lipopolysaccharide through down-regulating miR-146a. Int Immunopharmacol 69:373-381 (2019). Read more (PubMed: 30776646) »
  • Kano SI  et al. Glutathione S-transferases promote proinflammatory astrocyte-microglia communication during brain inflammation. Sci Signal 12:N/A (2019). Read more (PubMed: 30783009) »
See all 101 Publications for this product

ab1793 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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1-10 of 46 Abreviews or Q&A

Western blot abreview for Anti-TNF alpha antibody [52B83]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (C42B Prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
C42B Prostate cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More

Dr. Dimitra Kalamida

Verified customer

投稿 May 29 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-TNF alpha antibody [52B83]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (LNCaP C42B prostate cell line)
Permeabilization
Yes - 0.1% Triton X-100
Specification
LNCaP C42B prostate cell line
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Dr. Dimitra Kalamida

Verified customer

投稿 Jun 21 2018

Western blot abreview for Anti-TNF alpha antibody [52B83]

 Below Average
Abreviews
abreview image
Application
Western blot
Sample
Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

投稿 Jan 22 2013

Question

Hello,



I would like to investigate TNF-alpha expression rat perivascular adipose tissue using immunohistochemistry (paraffin sections) and western blotting. Do you know if ab1793 would be suitable to determine expression in rat PVAT or any other white adipose tissue? I haven’t been able to find any specific references in the literature.



Many thanks

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Answer

Thank you for your inquiry.

I am happy to confirm that ab1793 is tested and guaranteed for rat and WB and IHC-P.

I can confirm that we do not have any feedback about this antibody in perivascular adipose tissue yet. I therefore suggest running a positive control with a tissue that is known for its expression of TNF alpha.

I hope this information is helpful and wish you good luck with your experiments.

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Question

We see no staining for TNF alpha in sections of frozen PFA-fixed mouse brain and retina.

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Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

Dear all,
We had a look and ab1793 and ab18672 seem to look the most suitable. Would you be able to please e-mail him the epitopes of these monoclonals

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Answer

Thank you for contacting us.

Unfortunately there is no information available about the epitope recognised by these two antibodies, as it hasn’t been mapped.

I am very sorry I couldn’t help you further. Please do not hesitate to contact us if you need any other advice or information.

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Question

Thank you very much again, I am working on Human cell lines so I thinkab9739 will be good for our work.

Thank you so much.

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Answer

Thank you very much for your email.

My colleague is out of the office today but I will be happy to assist you with this issue.

I'm sending a free of charge vial of ab9739 on the order ***, which should arrive tomorrow. Please keep us updated about the results with this replacement antibody.

If you have any questions or need anything else, please let us know and we will be happy to help. Have a nice day.

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Question

Thank you very much again for your concern, yes this is our order.
So, if you can send me another good antibody for TNF-alpha it will be nice.

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Answer

Thank you for your reply. Are you working in any species or applications other than WB on human samples? We have a large number of antibodies to TNF alpha that are guaranteed for that purpose. For example, our rabbit polyclonal antibodies ab6671 and ab9739 have good customer feedback and have been used in a number of publications. Please let me know which antibody you would like to receive and I will be happy to process your replacement.

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Question

Thank you for your reply, and this is my protocol.



1. I transfected MDA cells in lane 2 and hTERT cells in lane 4 with the TNF-alpha gene. Lane 1:MDA-vector lane4:hTERT vector.

2. Lysed the cells with Ripa buffer.

3. 25 ug loading. DTT+loading dye.

4. Blocking by 5 % dry non fat milk with 1 % BSA in TBST.

5. Anti-TNF dil 1:1000. 4 C overnight.

6. Secondary Ab is 1:5000 anti mouse. 1 hr RT.



but I have no idea about PO number.

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Answer

Thank you for your reply. Based on the protocol you provided, I would not expect the bands observed on your blot to be specific. I was able to locate an order that was shipped to *** last week for ab1793 and ab15563. Is this the correct order? If I am able to locate your order, I would be happy to offer you a free of charge replacement, credit, or refund. Please let me know how you would like to proceed and I will be happy to help you further.

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Question

May I ask you a help,

Actually I have received the Anti-TNF alpha antibody [52B83] (ab1793) from you this week and I am trying to detect the TNF-alpha that is supposed to be overexpressed in some transfected cells, but after the W.B. I got strange bands around 40 KD and no bands at the position of 17 KD. I attched the fim after 5 min exposure.

Please, would you explain this to me.

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Answer

Thank you for contacting us. While TNF-alpha is known to form a homotrimer, the expected molecular weight for that band would be around 58 kDa. In order to help determine the nature of the bands you are observing, it would be helpful if you could please provide some additional details about your protocol.

1.) What type of cells were you staining, and what species was the transfected TNF alpha? Was the transfected protein tagged? What is the difference between the samples in lanes 1 / 2 and lanes 3 / 4?

2.) How were the cells lysed, how much protein was loaded per lane, and were the samples reduced and denatured?

3) What blocking agent was used?

4) What was the primary antibody dilution and incubation time?

5) What secondary antibody was used, what was the dilution and incubation time, and has this secondary antibody been working well with other primary antibodies?

6) What was your original order or PO number?

I will be happy to help you further once I have this additional information.

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1-10 of 46 Abreviews or Q&A

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