Recombinant
RabMAb

Recombinant Anti-TNF Receptor II antibody [EPR1653] (ab109322)

Overview

  • Product name

    Anti-TNF Receptor II antibody [EPR1653]
    See all TNF Receptor II primary antibodies
  • Description

    Rabbit monoclonal [EPR1653] to TNF Receptor II
  • Host species

    Rabbit
  • Specificity

    We do not guarantee IHC-P for mouse and rat species.
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human TNF Receptor II aa 400 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P20333
    (Peptide available as ab192471)

  • Positive control

    • WB: Jurkat, MCF7, SW480 and THP1 cell lysates IHC-P: uterus tissue
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109322 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 1/250 - 1/500.

For best results in ICC/IF please use 100% Methanol fixation.

WB 1/10000 - 1/50000. Detects a band of approximately 73 kDa (predicted molecular weight: 48 kDa).Can be blocked with TNF Receptor II peptide (ab192471).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250.

Antigen retrieval is recommended. We do not guarantee IHC-P for mouse and rat species.

Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, 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

  • All lanes : Anti-TNF Receptor II antibody [EPR1653] (ab109322) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : SW480 cell lysate
    Lane 4 : THP1 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 48 kDa
    Observed band size: 73 kDa
    why is the actual band size different from the predicted?

  • ab109322 staining TNF Receptor II in SW480 cells. The cells were fixed with 100% methanol. In our hands PFA fixed cells did not produce good results with this antibody. Nuclear DNA was labelled in blue with DAPI.

  • ab109322, at a 1/100 dilution, staining TNF Receptor II in paraffin embedded Human uterus tissue by Immunohistochemistry.

  • Overlay histogram showing HL60 cells stained with ab109322 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109322, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Primary: ab109322, 1/1000 dilution

    Sample: Jurkat whole cell lysate, 100 ug

    Secondary: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, 1/1000 dilution

    Capture antibody: 1:40 dilution (1µg in 1 mg lysate)

    Blocking and diluting buffer: 5% NFDM/TBST



    Anti-TNF Receptor II antibody [EPR1653] (ab109322) at 1/1000 dilution + Jurkat whole cell lysate at 100 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Observed band size: 73 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:

See all 9 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you very much for your reply and for your call today.

I'm sending a free of charge vial of ab7187 RAW 264.7 cell lysate, on the order ***. This lysate should arrive tomorrow.

Please keep me updated about the results with ab109322 and this lysate, and let me know if you have any further questions or need anything else. Have a nice evening!

Read More

Answer

Thank you again for your email, and for your patience.

I've heard back from the lab, and they load 10 ug of protein per lane. None of the cell lines are transfected, and they use the general Epitomics Western blot protocol-

http://www.epitomics.com/products/support/wb

Please let me know which positive control lysate you would like to try. Have a nice day.

Read More

Answer

Thank you very much for your reply.

I've asked the lab whether the cells are transfected, and how many micrograms of lysate were loaded.

I'd be happy to send a positive control lysate, however I can't send all of the lysates shown in the image. We have the RAW 264.7 lysate ab7187 or NIH/3T3 lysate ab7179-

https://www.abcam.com/RAW-264-7-Mouse-leukaemic-monocyte-macrophage-cell-line-Whole-Cell-Lysate-ab7187.html

https://www.abcam.com/NIH-3T3-Mouse-embryonic-fibroblast-cell-line-Whole-Cell-Lysate-ab7179.html

Which of these would you prefer? I'll be happy to send this out promptly. Please forward me your original PO or order number and I will ship it to the same address.

I look forward to hearing from you, and I will send you the information from the lab once I receive it. Please let me know if you have further questions.

Read More

Question
Answer

Thank you very much for your call yesterday.

I've received the attached image from the lab, which shows Western blot results using C6, Raw264.7, PC12, and NIH/3T3 cell lysates.

The band probably aligns most closely with the ˜ 60 kDa band that you described, but please let me know if testing with the alternative antibody ab119998 would be helpful, and I'll send this out to you.

I look forward to hearing from you. If you have further questions or need anything else, please let me know and I'll be happy to help.

Read More

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