Overview

  • Product name
    Anti-TNF Receptor II antibody [MR2-1]
    See all TNF Receptor II primary antibodies
  • Description
    Mouse monoclonal [MR2-1] to TNF Receptor II
  • Host species
    Mouse
  • Specificity
    The antibody reacts with the extra-cellular part of the TNF-R II and with the soluble receptor.
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Human, Cynomolgus monkey, Rhesus monkey
  • Immunogen

    soluble TNF-R75 from culture supernatant of cells transfected with the extracellular part of TNF-R75, purified by affinity chromatography on a TNF-coupled agarose column.

  • Positive control
    • Recombinant human Soluble TNF Receptor II protein (ab640) can be used as a positive control in WB.

Properties

Applications

Our Abpromise guarantee covers the use of ab8161 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use at an assay dependent concentration. Predicted molecular weight: 46 kDa.
IP Use at an assay dependent concentration.
Flow Cyt Use 2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Receptor with high affinity for TNFSF2/TNF-alpha and approximately 5-fold lower affinity for homotrimeric TNFSF1/lymphotoxin-alpha. The TRAF1/TRAF2 complex recruits the apoptotic suppressors BIRC2 and BIRC3 to TNFRSF1B/TNFR2. This receptor mediates most of the metabolic effects of TNF-alpha. Isoform 2 blocks TNF-alpha-induced apoptosis, which suggests that it regulates TNF-alpha function by antagonizing its biological activity.
  • Sequence similarities
    Contains 4 TNFR-Cys repeats.
  • Post-translational
    modifications
    Phosphorylated; mainly on serine residues and with a very low level on threonine residues.
    A soluble form (tumor necrosis factor binding protein 2) is produced from the membrane form by proteolytic processing.
  • Cellular localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD120b antibody
    • p75 antibody
    • p75 TNF receptor antibody
    • p75TNFR antibody
    • p80 TNF alpha receptor antibody
    • p80 TNF-alpha receptor antibody
    • Soluble TNFR1B variant 1 antibody
    • TBP-2 antibody
    • TBPII antibody
    • TNF R II antibody
    • TNF R2 antibody
    • TNF R75 antibody
    • TNF-R2 antibody
    • TNF-RII antibody
    • TNFBR antibody
    • TNFR-II antibody
    • TNFR1B antibody
    • TNFR2 antibody
    • TNFR80 antibody
    • TNFRII antibody
    • Tnfrsf1b antibody
    • TNR1B_HUMAN antibody
    • Tumor necrosis factor beta receptor antibody
    • Tumor necrosis factor binding protein 2 antibody
    • Tumor necrosis factor receptor 2 antibody
    • Tumor necrosis factor receptor superfamily member 1B antibody
    • Tumor necrosis factor receptor type II antibody
    • Tumor necrosis factor-binding protein 2 antibody
    see all

Images

  • ICC/IF image of ab8161 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8161, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • HDMECs were harvested with trypsin and washed three times with ice-cold PBS. Lysis buffer (50 mm Tris-HCl, pH 7.5, 10% of glycerol, 1% of Triton X-100, 300 mm NaCl, 150 mm KCl, 5 mm EDTA, 1 mm dithiothreitol, 0.5 mm Na3VO4, 10 mm NaF, 10 mm iodoacetamide, and protease inhibitors) or SDS gel buffer with ß-mercaptoethanol was then added, and the lysates were cleared of debris by centrifugation. 80 µg of total protein/sample was subjected to 4–15% gradient SDS-PAGE and electrophoretically transferred to polyvinylidene difluoride membranes. Protein blots on the membrane were identified by using ab8161 followed by horseradish peroxidase-conjugated anti-IgG secondary antibodies in blocking buffer. Images were obtained by chemiluminescence on x-ray film using ECL reagents.
  • Overlay histogram showing HL60 cells stained with ab8161 (red line). The cells were fixed with 80% 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 (ab8161, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Miettinen JA  et al. Tumor necrosis factor alpha promotes the expression of immunosuppressive proteins and enhances the cell growth in a human bone marrow-derived stem cell culture. Exp Cell Res 317:791-801 (2011). ICC/IF ; Human . Read more (PubMed: 21182837) »
  • Ding B  et al. FOXO3a regulates oxygen-responsive expression of tumor necrosis factor receptor 2 in human dermal microvascular endothelial cells. J Biol Chem 284:19331-9 (2009). WB ; Human . Read more (PubMed: 19473970) »
See all 4 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for contacting us. We may be able to custom make this antibody in an azide-free form for bulk orders of 10 or more vials. If you are interested in a custom bulk purchase, please contact my colleagues at mailto:sales@abcam.com and they can help put together a quote and estimated availability time for you.

Alternatively, for smaller purchases you could use one of our concentration and clean up kits (ab102778) to perform a buffer exchange and greatly reduce the concentration of azide in the antibody. I hope this helps, please let me know if you need any additional information or assistance.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (TF-1 cells)
Loading amount
30 µg
Specification
TF-1 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Aug 01 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (TF-1 cells)
Total protein in input
30 µg
Specification
TF-1 cells
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted Sep 18 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Brain microvascular endothelial cells)
Specification
Brain microvascular endothelial cells
Permeabilization
No

Abcam user community

Verified customer

Submitted Feb 02 2007

Answer

Thank you for your enquiry. We have just received the following information from the source of ab8161: The antibody to human CD120b, 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. This article shows 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. We hope this information will be helpful to you. If you have further questions please do not hesitate to contact us.

Read More

Answer

I'm afraid that the precise AA seq that this antibody recognises has not been determined.

Read More

Question
Answer

This antbody has not yet been tested on rat tissue

Read More

Question
Answer

The agonistic properties of HM2007 are tested in a cytotoxicity assay with KYM-1 cells. In this cytotoxicity assay concentrations of 0.03 ; 0.3 and 3 microgram/ml antibody were used.

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up