Recombinant
RabMAb

Recombinant Anti-TNFAIP3 antibody [EPR2663] - BSA and Azide free (ab227987)

Overview

  • Product name

    Anti-TNFAIP3 antibody [EPR2663] - BSA and Azide free
    See all TNFAIP3 primary antibodies
  • Description

    Rabbit monoclonal [EPR2663] to TNFAIP3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human TNFAIP3 aa 500-600.
    (Peptide available as ab175807)

  • Positive control

    • WB: Jurkat cells treated with TNF and TPA and Daudi cell lysates. IHC-P: Human kidney tissues. ICC/IF: Daudi and HeLa cells. Flow Cyt: HepG2 and Daudi cells.
  • General notes

    Ab227987 is the carrier-free version of ab92324. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227987 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227987 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 Use at an assay dependent concentration. Predicted molecular weight: 90 kDa.Can be blocked with TNFAIP3 peptide (ab175807).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Ubiquitin-editing enzyme that contains both ubiquitin ligase and deubiquitinase activities. Essential component of a ubiquitin-editing protein complex, comprising also RNF11, ITCH and TAX1BP1, that ensures the transient nature of inflammatory signaling pathways. Upon TNF stimulation, deubiquitinates 'Lys-63'-polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NF-kappa-B. In vitro able to deubiquitinate both 'Lys-48'- and 'Lys-63' polyubiquitin chains. Inhibitor of programmed cell death. Has a role in the function of the lymphoid system.
  • Sequence similarities

    Belongs to the peptidase C64 family.
    Contains 7 A20-type zinc fingers.
    Contains 1 OTU domain.
  • Domain

    The A20-type zinc fingers mediate the ubiquitin ligase activity.
    The OTU domain mediates the deubiquitinase activity.
  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • A20 antibody
    • AISBL antibody
    • MGC104522 antibody
    • MGC138687 antibody
    • MGC138688 antibody
    • OTU domain containing protein 7C antibody
    • OTU domain-containing protein 7C antibody
    • OTUD7C antibody
    • Putative DNA binding protein A20 antibody
    • Putative DNA-binding protein A20 antibody
    • TNAP3_HUMAN antibody
    • TNF alpha-induced protein 3 antibody
    • TNFA1P2 antibody
    • TNFAIP 3 antibody
    • TNFAIP3 (A20) antibody
    • TNFAIP3 antibody
    • Tumor necrosis factor alpha induced protein 3 antibody
    • Tumor necrosis factor alpha-induced protein 3 antibody
    • Tumor necrosis factor induced protein 3 antibody
    • Tumor necrosis factor inducible protein A20 antibody
    • tumor necrosis factor, alpha-induced protein 3 antibody
    • Zinc finger protein A20 antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling TNFAIP with purified ab92324 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Flow cytometry analysis of Daudi cells labelling TNFAIP3 with purified ab92324 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Flow cytometry analysis of Daudi cells labelling TNFAIP3 with unpurified ab92324 at 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Immunocytochemsitry/Immunofluorescence analysis of Daudi cells labelling TNFAIP3 (red) with purified ab92324 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Immunocytochemsitry/Immunofluorescence analysis of Daudi cells labelling TNFAIP3 (red) with unpurified ab92324 at 1/50. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with purified ab92324 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • ICC/IF image of unpurified ab92324 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab92324, neat) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • Overlay histogram showing HepG2 cells stained with unpurified ab92324 (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. The cells were then incubated with the antibody (ab92324, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92324).

  • This IHC data was generated using the same anti-TNFAIP3 antibody clone, EPR2663, in a different buffer formulation (cat# ab92324).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with unpurified ab92324 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

References

This product has been referenced in:

  • Kwon DJ  et al. Generation of a-1,3-galactosyltransferase knocked-out transgenic cloned pigs with knocked-in five human genes. Transgenic Res 26:153-163 (2017). WB . Read more (PubMed: 27554374) »
  • Ginster S  et al. Two Antagonistic MALT1 Auto-Cleavage Mechanisms Reveal a Role for TRAF6 to Unleash MALT1 Activation. PLoS One 12:e0169026 (2017). WB . Read more (PubMed: 28052131) »
See all 9 Publications for this product

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