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  1. Link

    tnf-alpha-antibody-ab66579.pdf

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Immunology Innate Immunity Cytokines TNF Superfamily
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Anti-TNF alpha antibody (ab66579)

  • Datasheet
  • SDS
Reviews (10)Q&A (11)References (102)

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Western blot - Anti-TNF alpha antibody (ab66579)
  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody (ab66579)

Key features and details

  • Rabbit polyclonal to TNF alpha
  • Suitable for: WB, ICC/IF
  • Reacts with: Mouse, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-TNF alpha antibody [EPR19147] (ab183218)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-TNF alpha antibody
    See all TNF alpha primary antibodies
  • Description

    Rabbit polyclonal to TNF alpha
  • Host species

    Rabbit
  • Specificity

    Ab66579 has been batch tested in Western Blot using LPS stimulated lysates. Abcam does not recommend this product for use in IHC. Please contact Abcam Scientific Support for more information.
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cat, Chimpanzee, Baboon
  • Immunogen

    Synthetic peptide corresponding to Human TNF alpha aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab66578)

  • Positive control

    • This antibody gave a positive signal in the following LPS stimulated lysates: THP1, RAW 264.7 ICC/IF: PMA/Befreldin/LPS treated RAW 264.7 cell line.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • TNF Superfamily
    • Signal Transduction
    • Growth Factors/Hormones
    • TNF
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Growth factors
    • TNF
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Metabolism
    • Types of disease
    • Metabolic disorders
    • Neuroscience
    • Development
    • Neuroscience
    • Diseases
    • Neuroscience
    • Processes

Associated products

  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human TNF alpha protein (Active) (ab259410)
    • Recombinant mouse TNF alpha protein (Active) (ab259411)
  • Related Products

    • Human TNF alpha ELISA Kit (ab181421)
    • Mouse TNF alpha ELISA Kit (ab208348)
    • Human TNF alpha Matched Antibody Pair Kit (ab213467)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab66579 in the following tested applications.

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

Application Abreviews Notes
WB (5)
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 25 kDa).

Abcam recommends using 3% Milk as the blocking agent.

ICC/IF (2)
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 26 kDa (predicted molecular weight: 25 kDa).

Abcam recommends using 3% Milk as the blocking agent.

ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    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.
  • Involvement in disease

    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).
  • Sequence similarities

    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications

    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.
  • Cellular localization

    Secreted and Cell membrane.
  • Target information above from: UniProt accession P01375 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7124 Human
    • Entrez Gene: 21926 Mouse
    • Entrez Gene: 24835 Rat
    • Omim: 191160 Human
    • SwissProt: P19101 Cat
    • SwissProt: Q8HZD9 Chimpanzee
    • SwissProt: P01375 Human
    • SwissProt: P06804 Mouse
    • SwissProt: P16599 Rat
    • Unigene: 241570 Human
    • Unigene: 1293 Mouse
    • Unigene: 2275 Rat
    see all
  • Alternative names

    • 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

Images

  • Western blot - Anti-TNF alpha antibody (ab66579)
    Western blot - Anti-TNF alpha antibody (ab66579)
    All lanes : Anti-TNF alpha antibody (ab66579) at 1 µg/ml

    Lane 1 : THP1 cells treated with PMA (negative control)
    Lane 2 : THP1 cells treated with PMA+LPS+Brefeldin A
    Lane 3 : Raw264.7 cells treated with PMA (negative control)
    Lane 4 : Raw264.7 cells treated with PMA+LPS+Brefeldin A

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?
    Additional bands at: 28 kDa (possible glycosylated form), 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 2 minutes


    Mature THP-1 and Raw264.7 cells were treated with LPS (1 ug/ml) for 6 hours to stimulate production of TNF alpha. Brefeldin A (300 ng/ml) was added in the final 3 hours of incubation to inhibit golgi traffic.

  • Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody (ab66579)
    Immunocytochemistry/ Immunofluorescence - Anti-TNF alpha antibody (ab66579)

    ab66579 staining TNF alpha in Raw264.7 cells. The cells were treated with LPS for 7 hours and Brefeldin A (ab120299) for the final 3 hours. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab66579 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

     

    Also suitable in cells fixed with 4% paraformaldehyde (10 min).

     

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (102)

Publishing research using ab66579? Please let us know so that we can cite the reference in this datasheet.

ab66579 has been referenced in 102 publications.

  • Li X  et al. miR-142-3p targets AC9 to regulate sciatic nerve injury-induced neuropathic pain by regulating the cAMP/AMPK signalling pathway. Int J Mol Med 47:561-572 (2021). PubMed: 33416140
  • Jin Q  et al. Allisartan isoproxil attenuates oxidative stress and inflammation through the SIRT1/Nrf2/NF-?B signalling pathway in diabetic cardiomyopathy rats. Mol Med Rep 23:N/A (2021). PubMed: 33495841
  • Parekh SV  et al. Hippocampal TNF-a Signaling Mediates Heroin Withdrawal-Enhanced Fear Learning and Withdrawal-Induced Weight Loss. Mol Neurobiol 58:2963-2973 (2021). PubMed: 33580871
  • Derbis M  et al. Short antisense oligonucleotides alleviate the pleiotropic toxicity of RNA harboring expanded CGG repeats. Nat Commun 12:1265 (2021). PubMed: 33627639
  • Zhang J  et al. Minocycline attenuation of rat corpus callosum abnormality mediated by low-dose lipopolysaccharide-induced microglia activation. J Neuroinflammation 18:100 (2021). PubMed: 33902641
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 21 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-TNF alpha antibody

Average
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Neuron)
Permeabilization
Yes - Permeabilized with 0.4% Triton X-100 in PBS for 1 hour at room temperature
Specification
Neuron
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 07 2021

Western blot abreview for Anti-TNF alpha antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HepG2 cells)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
35 µg
Specification
HepG2 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 06 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-TNF alpha antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mice hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Mice hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Jul 03 2020

Western blot abreview for Anti-TNF alpha antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mice hepatocytes)
Gel Running Conditions
Reduced Denaturing (4-15 % SDS-PAGE)
Loading amount
40 µg
Specification
Mice hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Jul 02 2020

Flow Cytometry abreview for Anti-TNF alpha antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Flow Cytometry
Sample
Rat Cell (Brain)
Permeabilization
Yes - 0.25% Triton X-100
Gating Strategy
a single cells population
Specification
Brain
Fixation
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MS. Aneta Radziwon-Balicka

Verified customer

Submitted Sep 13 2017

Western blot abreview for Anti-TNF alpha antibody

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Placenta)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Placenta
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Vicky Valued Customer

Verified customer

Submitted Dec 22 2016

Western blot abreview for Anti-TNF alpha antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (LIVER)
Gel Running Conditions
Reduced Denaturing
Loading amount
50 µg
Specification
LIVER
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 06 2016

Question

We work with immunohistochemistry on mouse knee joints. The suggested antibody-ab71281 doesn’t seem to work with mouse tissue. But I found ab66579 antibody which seems to work on mouse tissue.

I want to know if this antibody(ab66579- immunogen-human TNF-alpha) has strong affinity for the mouse tissue as ab34674(immunogen-mouse TNF-alpha).

Regards

Read More

Abcam community

Verified customer

Asked on May 23 2013

Answer

Thank you for contacting us.

The antibodies ab66579 and ab71281 are polyclonal antibodies and are guaranteed form detection of mouse TNF alpha protein. We do not determine the affinity of polyclonal antibodies which is rather difficult because polyclonal antibodies are a mixture a of high affinity and low affinity IgGs.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on May 23 2013

Question

Name: Markus Brückner
Product code: 17611

Inquiry: Dear Sirs, I´m searching for a biotinylated anti-mouse mAB against the membrane form of TNF-a, rather not the solulable form of TNF-a. Is the above mentioned product right for this? Or is there a better biotinylated mAb against membrane TNF-a? Thanks alot for your support.

Read More

Abcam community

Verified customer

Asked on Dec 13 2012

Answer

Thank you for your enquiry.

I am sorry to confirm that most of our TNF alpha antibodies, including ab17611, have an epitope within the soluble region of the protein.

I have found one antibody, tested in mouse, that has an immunogen within the membrane region only. As far as I am aware, the soluble form is from amino acids 80-235. This antibody has an immunogen within the first 50 amino acids of the protein so should detect membrane bound TNF alpha only. This region is cytoplasmic.

TNF alpha antibody (ab66579)
Tested in: IHC-Fr, WB, IHC-P
Species reactivity: Mouse, Rat, Human
https://www.abcam.com/TNF-alpha-antibody-ab66579.html (or use the following: https://www.abcam.com/TNF-alpha-antibody-ab66579.html).

I am sorry this is not biotinylated. However, we do have some Easylink conjugation kits available including some for biotinylation of primary antibodies. I can recommend to review the details on the online datasheet:

ab102864 Biotin conjugation kit
https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-100g-Biotin-ab102864.html (or use the following: https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-100g-Biotin-ab102864.html).

Because of the high percentage of BSA in the ab66579 antibody, I can recommend to also consider using our Concentration and Purification Kits (https://www.abcam.com/antibody-concentration-kit-3-columns-ab102778.html or https://www.abcam.com/antibody-purification-kit-3-purifications-ab102784.html) before conjugation to ensure the conjugation will work.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Dec 13 2012

Question

Thank you for the response.
please the antibody here are they for both ELISA and Immunohistochemistry or for one only.
I tried sending you email the other day but it was not going.
Thank you



Read More

Abcam community

Verified customer

Asked on Oct 30 2012

Answer

Please check the online datasheets of all antibodies. You will find the required information.

Thanks!

Read More

Abcam Scientific Support

Answered on Oct 30 2012

1-10 of 21 Abreviews or Q&A

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