Recombinant
RabMAb

Recombinant Anti-SIGIRR antibody [EPR22438-93] (ab228977)

Overview

  • Product name

    Anti-SIGIRR antibody [EPR22438-93]
    See all SIGIRR primary antibodies
  • Description

    Rabbit monoclonal [EPR22438-93] to SIGIRR
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Mouse SIGIRR aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9JLZ8

  • Positive control

    • IHC-P: Human kidney and colon tissue; Mouse kidney tissue; Rat kidney tissue WB: Hepa1-6 cell lysate; Mouse stomach, kidney, spleen and pancreas tissue lysate; Human stomach and fetal kidney lysate; Rat stomach and liver tissue lysate Flow cyt: MEF and Hepa1-6 cells IP: Hepa1-6 whole cell lysate
  • 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

  • Form

    Liquid
  • Storage instructions

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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22438-93
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab228977 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
Flow Cyt 1/600.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 46 kDa).
IP 1/30.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      Acts as a negative regulator of the Toll-like and IL-1R receptor signaling pathways. Attenuates the recruitment of receptor-proximal signaling components to the TLR4 receptor, probably through an TIR-TIR domain interaction with TLR4. Through its extracellular domain interferes with the heterodimerization of Il1R1 and IL1RAP.
    • Tissue specificity

      Mainly expressed in epithelial tissues such as kidney, lung and gut.
    • Sequence similarities

      Belongs to the interleukin-1 receptor family.
      Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
      Contains 1 TIR domain.
    • Cellular localization

      Membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • Ig IL1 related receptor antibody
      • MGC110992 antibody
      • SIGIR_HUMAN antibody
      • SIGIRR antibody
      • Single Ig IL 1 related receptor antibody
      • Single Ig IL-1-related receptor antibody
      • Single Ig IL-1R-related molecule antibody
      • Single Ig IL1R Related Molecule antibody
      • Single immunoglobulin and toll interleukin 1 receptor (TIR) domain antibody
      • Single immunoglobulin and toll interleukin 1 receptor domain antibody
      • Single immunoglobulin domain containing IL1R related protein antibody
      • Single immunoglobulin domain IL1R1 related antibody
      • Single immunoglobulin domain-containing IL1R-related protein antibody
      • TIR 8 antibody
      • TIR8 antibody
      • Toll/interleukin 1 receptor 8 antibody
      • Toll/interleukin-1 receptor 8 antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney (PMID:14715412, 12925853). The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized MEF (mouse embryonic fibroblast (immortalized)) (Left) / Hepa1-6 (mouse hepatoma epithelial cell line) (Right) cells labelling SIGIRR with ab228977 at 1/600 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

      Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

      Negative control: MEF (PMID 12925853).

    • IGFBP5 was immunoprecipitated from 0.35 mg Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate 10μg with ab254324 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab228977 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

      Lane 1: Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate 10μg

      Lane 2: ab254324 IP in Hepa1-6 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228977 in Hepa1-6 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 5 seconds.

    • Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution  followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse kidney (PMID:14715412, 12925853). The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SIGIRR with ab228977 at 1/1000 dilution  followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon (PMID:21077278) The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SIGIRR with ab228977 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney (PMID:14715412, 12925853) The section was incubated with ab228977 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • All lanes : Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

      Lane 1 : Mouse kidney tissue lysate
      Lane 2 : Mouse kidney treated lysate treated with with PNGase F

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 46 kDa
      Observed band size: 46, 80 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature (PMID: 26344057).

      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

      Exposure time: 92 seconds.

    • All lanes : Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

      Lane 1 : Human fetal kidney tissue lysate
      Lane 2 : Rat liver tissue lysate
      Lane 3 : Mouse kidney tissue lysate
      Lane 4 : Mouse spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody (HRP))
      Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 46 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      The different apparent molecular weights are due to different levels of glycosylation. (PMID: 26344057).

      Exposure times.
      Lane 1: 48 seconds; Lane 2: 7.75 seconds; Lane 3: 70 seconds; Lane 4: 3 minutes.

      Blocking/diluting buffer and concentration: 5% NFDM/TBST.

    • All lanes : Anti-SIGIRR antibody [EPR22438-93] (ab228977) at 1/1000 dilution

      Lanes 1 & 5 : Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate
      Lane 2 : MEF (immortalized mouse embryonic fibroblasts), whole cell lysate
      Lane 3 : Mouse stomach tissue lysate
      Lane 4 : Mouse pancreas tissue lysate
      Lane 6 : Rat stomach tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lanes 1-4 & 6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
      Lane 5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Human stomach tissue lysate)

      Predicted band size: 46 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      The different apparent molecular weights are due to different levels of glycosylation. (PMID: 26344057).

      Negative control: MEF (PMID 12925853)

      Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

      Exposure times.

      Lane 1: 15 seconds; Lanes 2-6: 3 minutes.

    References

    ab228977 has not yet been referenced specifically in any publications.

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