Overview

  • Product name

    Anti-Iba1 antibody [EPR16588]
    See all Iba1 primary antibodies
  • Description

    Rabbit monoclonal [EPR16588] to Iba1
  • Host species

    Rabbit
  • Specificity

    For ab178846 Abcam recommends blocking in 3% milk for cleanest results in WB. Blocking with BSA gives slightly higher background.
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Mouse Iba1 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: O70200

  • Positive control

    • WB: HL-60, THP-1, U937, RAW 264.7 and NR8383 whole cell lysates; Human, mouse and rat spleen lysates; Mouse testis and liver lysates. IHC-P: Human Cerebral cortex, human hippocampus; Rat and mouse normal brain tissues. Flow Cyt: U937 cells. IHC-Fr: Mouse Cerebral cortex 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 ab178846 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 1/160.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/500 - 1/2000. Detects a band of approximately 10, 15 kDa (predicted molecular weight: 17 kDa).

Abcam recommends blocking in 3% milk for cleanest results in WB. Blocking with BSA gives slightly higher background.

IHC-FoFr 1/200.
ICC/IF 1/500.

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

  • 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

  • 4% paraformaldehyde-fixed, 0.1% Triton permeabilized Mouse Cerebral cortex tissue labeling Iba1 (Green) using ab178846 at 1/200 dilution in immunohistochemical analysis. A Goat anti-Rabbit IgG (H+L) Alexa Fluor® 488 was used as the secondary antibody.

    Heat-induced antigen retrieval with Citrate buffer (10 mM Citric acid, 0.05% Tween 20, pH 6.0), at 90ºC for 15 min, using a water bath. Image: Iba1 ab178846 (green) and Hoechst (blue). Counterstained with Hoechst, 2 ug/ml for 5 min at room temperature. Mounted in Fluoromount-G.

  • Formaldehyde-fixed, paraffin-embedded cynomolgus monkey brain tissue stained for Iba1 using ab178846 at 1/6000 dilution in immunohistochemical analysis.

    Antigen Retrieval: Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA

  • All lanes : Anti-Iba1 antibody [EPR16588] (ab178846)

    Lane 1 : Mouse testis
    Lane 2 : Mouse liver
    Lane 3 : Rat testis
    Lane 4 : Rat liver

    Predicted band size: 17 kDa

  • IHC image of Iba1 staining in rat normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Flow cytometry analysis of 2% paraformaldehyde fixed U937 (human histiocytic lymphoma cell line) cells labeling Iba1 with ab178846 at 1/160 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

  • IHC image of Iba1 staining in mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Iba1 antibody [EPR16588] (ab178846) at 1/500 dilution

    Lane 1 : Human Iba1 recombinant protein at 0.1 µg
    Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
    Lane 3 : A431 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg
    Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate at 30 µg
    Lane 5 : Human spleen tissue lysate at 20 µg
    Lane 6 : Mouse spleen tissue lysate at 30 µg
    Lane 7 : Rat spleen tissue lysate at 30 µg
    Lane 8 : U937 (human histiocytic lymphoma cell line) whole cell lysate at 30 µg
    Lane 9 : MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate at 20 µg
    Lane 10 : THP-1 (human monocytic leukemia cell line) whole cell lysate at 30 µg
    Lane 11 : THP-1 whole cell lysate, PMA treated at 30 µg
    Lane 12 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 30 µg
    Lane 13 : C6 (rat glial tumor cell line) whole cell lysate at 30 µg
    Lane 14 : NR8383 whole cell lysate at 30 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17 kDa


    Exposure time: 1 minute


    Abcam recommends blocking in milk for cleaner blots with reduced background, in comparison to BSA. 

    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab178846 (anti-Iba1 antibody; 1/500 dilution) for 18 hours at 4°C. Antibody binding was detected using ab97040 (HRP-labelled goat anti-mouse IgG) at  1:50,000 dilution for 1 hour at room temperature and visualised using ECL development solution ab133406.

  • 0.1% Triton-X 100 permeabilized paraformaldehyde-fixed Mouse cell Microglia cells labeling Iba1 (green) using ab178846 at 1/500 dilution in ICC/IF analysis.

  • IHC image of Iba1 staining in human normal hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Iba1 antibody [EPR16588] (ab178846) at 1/10000 dilution

    Lane 1 : THP-1 (human monocytic leukemia cell line) whole cell lysate
    Lane 2 : U937 (human histiocytic lymphoma cell line) whole cell lysate
    Lane 3 : Human spleen whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 17 kDa
    Observed band size: 15 kDa
    why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM /TBST

  • Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Iba1 with ab178846 at a 1/2000 dilution showing cytoplasm and nuclear staining on Glial cells. Counter stained with hematoxylin. Prediluted HRP Polymer for Rabbit/Mouse IgG was used as the secondary aantibody. Negative control also shown.

     

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-Iba1 antibody [EPR16588] (ab178846) at 1/2000 dilution + HL-60 (human promyelocytic leukemia cell line) whole cell lysate at 10 µg

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

    Predicted band size: 17 kDa
    Observed band size: 10, 15 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration:

    5% NFDM /TBST.

    Based on sequence analysis, ab178846 recognizes 2 isoforms with the predicted MWs of 17KDa and 11KDa, respectively.

References

This product has been referenced in:

  • Yuan C  et al. OAB-14, a bexarotene derivative, improves Alzheimer's disease-related pathologies and cognitive impairments by increasing ß-amyloid clearance in APP/PS1 mice. Biochim Biophys Acta Mol Basis Dis 1865:161-180 (2019). Read more (PubMed: 30389579) »
  • Chauhan G  et al. Distinct influence of COX-1 and COX-2 on neuroinflammatory response and associated cognitive deficits during high altitude hypoxia. Neuropharmacology 146:138-148 (2019). Read more (PubMed: 30476507) »
See all 31 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.1% TX-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 19 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ab93684 Tris EDTA pH9.0
Permeabilization
Yes - 0.05% tween 20
Specification
Brain
Blocking step
sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Formaldehyde

Mr. David Ivancic

Verified customer

Submitted Jun 19 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Brain)
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Collin Gagne

Verified customer

Submitted May 29 2019

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Adult Brain section (Cortex))
Antigen retrieval step
None
Permeabilization
Yes - Triton-X100 0.3%
Specification
Adult Brain section (Cortex)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 22°C
Fixative
Paraformaldehyde

Miss. Helene Hirbec

Verified customer

Submitted Jan 25 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (cerebrum)
Permeabilization
Yes - 0.1% TritonX100 / PBS
Specification
cerebrum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Weihua Wang​

Verified customer

Submitted Sep 28 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (cerebellum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Permeabilization
No
Specification
cerebellum
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 19 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Cerebral cortex)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM Citrate buffer, pH 6.0
Permeabilization
Yes - 0.1% Triton in PBS
Specification
Cerebral cortex
Blocking step
1% BSA + 5% Serum + 0.3 M Glycine as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Sergi Bayod

Verified customer

Submitted Sep 06 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cynomolgus monkey Tissue sections (Brain, Cerebrum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Ventana CC1 high pH
Permeabilization
No
Specification
Brain, Cerebrum
Fixative
Formaldehyde

David Weil

Verified customer

Submitted Sep 05 2018

Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Aug 30 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Dog Tissue sections (Brain)
Permeabilization
Yes - Tween20
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 06 2018

1-10 of 22 Abreviews or Q&A

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