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Recombinant

Recombinant Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (ab283319)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Western blot - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (ab283319)
  • Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
  • Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [EPR16589] to Iba1 - Mouse IgG1
  • Suitable for: WB, IP, Flow Cyt (Intra), IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Iba1 antibody [EPR16589] - Mouse IgG1
    See all Iba1 primary antibodies

  • Description

    Mouse monoclonal [EPR16589] to Iba1 - Mouse IgG1

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IP, Flow Cyt (Intra), IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control

    • WB: THP-1 whole cell lysate. IHC-P: Human cerebrum, Human endometrium and Mouse cerebrum, Rat cerebrum tissues. ICC: THP-1 cells. Flow Cyt: THP-1 cell. IP: THP-1 cell.

  • General notes

    This mouse antibody has been engineered from a RabMAb parent antibody (ab178847). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283319 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
1/1000. Predicted molecular weight: 16 kDa.
IP
1/30.
Flow Cyt (Intra)
1/10000.
IHC-P
1/10000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
1/100.
Notes
WB
1/1000. Predicted molecular weight: 16 kDa.
IP
1/30.
Flow Cyt (Intra)
1/10000.
IHC-P
1/10000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.

Target

  • Function

    Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.

  • Tissue specificity

    Detected in T-lymphocytes and peripheral blood mononuclear cells.

  • Sequence similarities

    Contains 2 EF-hand domains.

  • Post-translational
    modifications

    Phosphorylated on serine residues.

  • Cellular localization

    Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AIF 1 antibody
    • AIF-1 antibody
    • Aif1 antibody
    • AIF1 protein antibody
    • AIF1_HUMAN antibody
    • Allograft inflammatory factor 1 antibody
    • Allograft inflammatory factor 1 splice variant G antibody
    • allograft inflammatory factor-1 splice variant Hara-1 antibody
    • balloon angioplasty responsive transcription antibody
    • BART 1 antibody
    • G1 antibody
    • G1 putative splice variant of allograft inflamatory factor 1 antibody
    • IBA 1 antibody
    • IBA1 antibody
    • interferon gamma responsive transcript antibody
    • Interferon responsive transcript 1 antibody
    • interferon responsive transcript factor 1 antibody
    • Ionized calcium binding adapter molecule 1 antibody
    • Ionized calcium-binding adapter molecule 1 antibody
    • ionized calcium-binding adapter molecule antibody
    • IRT 1 antibody
    • IRT1 antibody
    • Microglia response factor antibody
    • MRF1 antibody
    • Protein g1 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Iba1 with ab283319 at 1/10000 (0.114 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the microglia in rat cerebrum. The section was incubated with ab283319 for 10 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Western blot - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Western blot - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    All lanes : Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (ab283319) at 1/1000 dilution

    Lane 1 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
    Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 16 kDa
    Observed band size: 16 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

    Negative control: MCF7 (HPA database) 

    Exposure time: 136 seconds

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Iba1 with ab283319 at 1/10000 (0.114 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the microglia in human cerebrum. The section was incubated with ab283319 for 10 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling Iba1 with ab283319 at 1/10000 (0.114 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the immune cells in human endometrium. The section was incubated with ab283319 for 10 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Iba1 with ab283319 at 1/10000 (0.114 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the microglia in mouse cerebrum. The section was incubated with ab283319 for 10 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

  • Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 cells labelling Iba1 with ab283319 at 1/100 (11.36 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Confocal image showing cytoplasmic staining in THP-1 cells.

    Negative control: MCF7 

    Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (ab283319)
    Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (ab283319)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell, Left) / THP-1 (Human monocytic leukemia monocyte, Right) cells labelling Iba1 with ab283319 at 1/10000 dilution (0.01ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

    Negative control: MCF7.

  • Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

    Iba1 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte), whole cell lysate 10 ug with ab283319 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283319 at 1/1000 dilution. mouse IgG for IP (HRP) (ab131368) was used at 1/10000 dilution.

    Lane 1: THP-1 (human monocytic leukemia monocyte), whole cell lysate 10 ug

    Lane 2: ab283319 IP in THP-1 whole cell lysate

    Lane 3: Mouse monoclonal IgG (ab18443) instead of ab283319 in THP-1 whole cell lysate

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

    Exposure time: 32 seconds

     

  • Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)
    Anti-Iba1 antibody [EPR16589] - Chimeric (ab283319)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab283319? Please let us know so that we can cite the reference in this datasheet.

ab283319 has not yet been referenced specifically in any publications.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Iba1 antibody [EPR16589] - Mouse IgG1

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Deer Cell (Brain tissue)
Permeabilization
Yes - 1% Triton X-100 in PBS
Specification
Brain tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Jan 04 2022

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