Recombinant
RabMAb

Recombinant Anti-IL-1RA antibody [EPR6483] - BSA and Azide free (ab226101)

Overview

  • Product name

    Anti-IL-1RA antibody [EPR6483] - BSA and Azide free
    See all IL-1RA primary antibodies
  • Description

    Rabbit monoclonal [EPR6483] to IL-1RA - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: A431, 293T, MOLT4, HeLa, RAW264.7, NIH/3T3, and L6 cell lysates. IHC-P: Human, mouse and rat kidney tissues. ICC/IF: HeLa cells. Flow Cyt: A431 and HeLa cells. IP: NIH/3T3 cell lysate.
  • General notes

    Ab226101 is the carrier-free version of ab124962. 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.

    ab226101 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

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.
  • Dissociation constant (KD)

    KD = 8.40 x 10 -12 M
    Learn more about KD
  • Storage buffer

    pH: 7.40
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6483
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab226101 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. Detects a band of approximately 20 kDa (predicted molecular weight: 20 kDa).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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.
ELISA Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function

    Inhibits the activity of interleukin-1 by binding to receptor IL1R1 and preventing its association with the coreceptor IL1RAP for signaling. Has no interleukin-1 like activity. Binds functional interleukin-1 receptor IL1R1 with greater affinity than decoy receptor IL1R2; however, the physiological relevance of the latter association is unsure.
  • Tissue specificity

    The intracellular form of IL1RN is predominantly expressed in epithelial cells.
  • Involvement in disease

    Microvascular complications of diabetes 4
    Interleukin 1 receptor antagonist deficiency
  • Cellular localization

    Cytoplasm and Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • DIRA antibody
    • F630041P17Rik antibody
    • ICIL 1RA antibody
    • ICIL-1RA antibody
    • ICIL1RA antibody
    • IL-1ra antibody
    • IL-1ra3 antibody
    • IL-1RN antibody
    • IL1 inhibitor antibody
    • IL1F3 antibody
    • IL1RA antibody
    • IL1RA_HUMAN antibody
    • IL1RN (IL1F3) antibody
    • IL1RN antibody
    • Interleukin 1 receptor antagonist antibody
    • Interleukin-1 receptor antagonist protein antibody
    • Intracellular IL 1 receptor antagonist type II antibody
    • Intracellular interleukin 1 receptor antagonist (icIL 1ra) antibody
    • IRAP antibody
    • MGC10430 antibody
    • MVCD4 antibody
    • Type II interleukin 1 receptor antagonist antibody
    see all

Images

  • Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab124962 at a dilution of 1 in 20 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling IL1RA with purified ab124962 at a dilution of 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary 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 (ab124962).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling IL1RA with purified ab124962 at a dilution of 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary 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 (ab124962).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling IL1RA with purified ab124962 at a dilution of 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary 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 (ab124962).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling IL1RA with purified ab124962 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • Flow Cytometry analysis of HeLa cells labelling IL1RA with purified ab124962 at 1/20 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • ab124962 (purified) at 1/20 immunoprecipitating IL1RA in NIH/3T3 whole cell lysate.

    Lane 1 (input): NIH/3T3 whole cell lysate (10µg)

    Lane 2 (+): ab124962 + NIH/3T3 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124962 in NIH/3T3 whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • Overlay histogram showing A431 cells stained with unpurified ab124962 (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 (unpurified ab124962, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124962).

  • This IHC data was generated using the same anti-IL-1RA antibody clone, EPR6483, in a different buffer formulation (cat# ab124962).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling IL-1RA with unpurified ab124962 at a dilution of 1/100.

  • This ICC/IF data was generated using the same anti-IL-1RA antibody clone, EPR6483, in a different buffer formulation (cat# ab124962).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling IL-1RA with unpurified ab124962 at a dilution of 1/100.

References

ab226101 has not yet been referenced specifically in any publications.

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