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    iba1-antibody-ab5076.pdf

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Anti-Iba1 antibody (ab5076)

  • Datasheet
  • SDS
Reviews (90)Q&A (25)References (571)

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Western blot - Anti-Iba1 antibody (ab5076)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)

Key features and details

  • Goat polyclonal to Iba1
  • Suitable for: IHC-P, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Iba1 antibody [EPR16588] (ab178846)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Iba1 antibody
    See all Iba1 primary antibodies
  • Description

    Goat polyclonal to Iba1
  • Host species

    Goat
  • Specificity

    ab5076 is expected to recognize the isoforms represented by NP_001614 and NP_116573 but not NP_004838. 

     

  • Tested Applications & Species

    Application Species
    IHC-P
    Rat
    Human
    WB
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Iba1 aa 135-147 (C terminal). Accession Number(s): NP_116573.1; NP_001614.3
    Sequence:

    C-TGPPAKKAISELP


    Database link: P55008
    (Peptide available as ab23067)
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: Human lymph node and rat brain tissue lysate. IHC-P: Human placenta tissue.
  • General notes

    The Iba1 antibody (ab5076) is commonly used as a marker of microglia activation in staining and immunohistochemistry, given that ionized calcium binding adaptor molecule 1 (Iba1) is a microglia/macrophage-specific calcium-binding protein with actin-bundling activity that participates in membrane ruffling and phagocytosis in activated microglia.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.30
    Preservative: 0.02% Sodium azide
    Constituents: Tris buffered saline, 0.5% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Immunology
    • Immune Interventions
    • Transplantation
    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Neuroscience
    • Cell Type Marker
    • Glia marker
    • Microglia marker
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Signal Transduction
    • Antibodies
    • iba1

Associated products

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant Human Iba1 protein (ab105593)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-TMEM119 antibody [28-3] - Microglial marker (ab209064)
    • Anti-CD68 antibody [FA-11] (ab53444)
    • Anti-CD11b antibody [M1/70] (ab8878)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5076 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Rat
Human
WB
Rat
Human
All applications
Pig
Macaque monkey
Application Abreviews Notes
IHC-P (27)
Use a concentration of 6 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (7)
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 16-17 kDa (predicted molecular weight: 17 kDa).

A 1 hour primary incubation is recommended for this product. We received several excellent Abreviews on this antibody working with mouse, however, mouse is not a batch tested species and we cannot guarantee that this antibody will work on mouse. Some of our customers have observed high background in mouse samples. We recommend blocking in milk (3% milk in TBST) for 1 hour at room temperature. Blocking with BSA gives high background.

Notes
IHC-P
Use a concentration of 6 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 16-17 kDa (predicted molecular weight: 17 kDa).

A 1 hour primary incubation is recommended for this product. We received several excellent Abreviews on this antibody working with mouse, however, mouse is not a batch tested species and we cannot guarantee that this antibody will work on mouse. Some of our customers have observed high background in mouse samples. We recommend blocking in milk (3% milk in TBST) for 1 hour at room temperature. Blocking with BSA gives high background.

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.
  • Target information above from: UniProt accession P55008 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 199 Human
    • Entrez Gene: 397271 Pig
    • Entrez Gene: 29427 Rat
    • Omim: 601833 Human
    • SwissProt: P55008 Human
    • SwissProt: P81076 Pig
    • SwissProt: P55009 Rat
    • Unigene: 76364 Human
    • Unigene: 32080 Rat
    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

  • Western blot - Anti-Iba1 antibody (ab5076)
    Western blot - Anti-Iba1 antibody (ab5076)
    All lanes : Anti-Iba1 antibody (ab5076) at 1 µg/ml

    Lane 1 : Human lymph node tissue lysate in RIPA buffer
    Lane 2 : Rat brain tissue lysate in RIPA buffer

    Lysates/proteins at 35 µg per lane.

    Developed using the ECL technique.

    Predicted band size: 17 kDa



    Detected by chemiluminescence.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)

    Immunohistochemistry analysis of paraffin-embedded human placenta tissue labeling Iba1 with ab5076 at 6 µg/mL. Heat induced antigen retrieval with citrate buffer pH 6, HRP-staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)Image from Iris Margalit Trutzer et al, ACTA neuropathologica communications, 7, 40, Fig. 7; doi: 10.1186/s40478-019-0684-8

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human lateral prefrontal cortex tissue labeling iba1 labeled microglia (labeled with black arrows) within the cortex using ab5076 at 1/1000 dilution. (Cropped image).

    Sections were rinsed in 0.01 m PBS, pH 7.4, followed by 10% serum matching the species of the secondary antibody, 5% bovine serum albumin, and 0.1% Triton X-100 in 0.01 m PBS blocking solution for 1 h and incubated for 2 days at 4 °C in primary antibody. The sections were rinsed in PBS and incubated overnight at 4 °C with a horse anti-Goat IgG (Biotinylated) secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody (ab5076)This image is courtesy of an abreview submitted by Carl Hobbs (King's College London, United Kingdom)

    IHC-P image of Iba1 staining on rat brain sections using ab5076 (1:1000). The sections were deparaffinized and subjected to heat mediated antigen retrieval using citric acid. The sections were blocked using 1% BSA for 10 mins at 21°C. ab5076 was diluted 1:1000 and incubated with the sections for 2 hours at 21°C. The secondary antibody used was Rabbit polyclonal to anti sheep/goat IgG conjugated to biotin (1:200)

    See Abreview

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (571)

    Publishing research using ab5076? Please let us know so that we can cite the reference in this datasheet.

    ab5076 has been referenced in 571 publications.

    • Keane L  et al. mTOR-dependent translation amplifies microglia priming in aging mice. J Clin Invest 131:N/A (2021). IHC-P ; Mouse . PubMed: 33108356
    • Cai L  et al. Glioblastoma Exhibits Inter-Individual Heterogeneity of TSPO and LAT1 Expression in Neoplastic and Parenchymal Cells. Int J Mol Sci 21:N/A (2020). PubMed: 31963507
    • Zhu Y  et al. Single-Cell Analysis for Glycogen Localization and Metabolism in Cultured Astrocytes. Cell Mol Neurobiol 40:801-812 (2020). PubMed: 31863221
    • Xu AL  et al. Characterization of astrocytes and microglial cells in the hippocampal CA1 region after transient focal cerebral ischemia in rats treated with Ilexonin A. Neural Regen Res 15:78-85 (2020). PubMed: 31535655
    • Li L  et al. Foam cells promote atherosclerosis progression by releasing CXCL12. Biosci Rep 40:N/A (2020). PubMed: 31894855
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 115 Abreviews or Q&A

    Immunohistochemistry free floating abreview for Anti-Iba1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 24 2021

    Immunohistochemistry free floating abreview for Anti-Iba1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Cerebellum)
    Specification
    Cerebellum
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 24 2021

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Iba1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Cerebellum)
    Antigen retrieval step
    None
    Permeabilization
    Yes - PBS 1x, Triton x-100 1%
    Specification
    Cerebellum
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 18 2021

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Iba1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    None
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 18 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain (thalamus))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6.4
    Permeabilization
    No
    Specification
    Brain (thalamus)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 24°C
    Fixative
    Formalin
    Read More

    Mr. Yazead Buhidma

    Verified customer

    Submitted Sep 29 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 23°C
    Fixative
    Formaldehyde
    Read More

    Mr. Victor Bernal

    Verified customer

    Submitted Aug 26 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Iba1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (brain)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    brain
    Blocking step
    TBS, 0.5% Triton, 0.3 M glycine, 1% BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted May 26 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Iba1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (iPSC-derived Microglia)
    Permeabilization
    Yes - 0.1% triton 5min
    Specification
    iPSC-derived Microglia
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Boyd Kenkhuis

    Verified customer

    Submitted May 06 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Iba1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0, Water bath 20min 90 degrees
    Permeabilization
    Yes - PBS Tween
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 22 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Iba1 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (iPSC-derived microglia)
    Permeabilization
    Yes - PBS-1% Triton X-100
    Specification
    iPSC-derived microglia
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 18 2019

    1-10 of 115 Abreviews or Q&A

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