Anti-Iba1 antibody (ab153696)
Key features and details
- Rabbit polyclonal to Iba1
- Suitable for: ICC/IF, IHC-P, IP, WB, Flow Cyt
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- 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
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Product name
Anti-Iba1 antibody
See all Iba1 primary antibodies -
Description
Rabbit polyclonal to Iba1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, IP, WB, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment corresponding to Human Iba1 (internal sequence).
Database link: P55008 -
Positive control
- WB: Rat and mouse liver extract, THP-1, mouse and rat brain tissue; IHC-P: Rat and mouse brain tissue; Flow: human THP1 cells, murine microglia cells; ICC/IF: human THP-1 cells; IP: human K562 cells
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 79% PBS, 20% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab153696 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 |
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ICC/IF |
1/100 - 1/1000.
Not suitable for mouse |
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IHC-P |
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
1/100 - 1/500.
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WB |
1/500 - 1/10000. Predicted molecular weight: 16 kDa.
For ab153696 Abcam recommends blocking in 5% milk for cleanest results in WB. Blocking with BSA gives slightly higher background. |
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Flow Cyt |
1/50 - 1/200.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
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Notes |
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ICC/IF
1/100 - 1/1000. Not suitable for mouse |
IHC-P
1/100 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/100 - 1/500. |
WB
1/500 - 1/10000. Predicted molecular weight: 16 kDa. For ab153696 Abcam recommends blocking in 5% milk for cleanest results in WB. Blocking with BSA gives slightly higher background. |
Flow Cyt
1/50 - 1/200.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
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Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 199 Human
- Entrez Gene: 11629 Mouse
- Entrez Gene: 29427 Rat
- Omim: 601833 Human
- SwissProt: P55008 Human
- SwissProt: O70200 Mouse
- SwissProt: P55009 Rat
- Unigene: 76364 Human
see all -
Alternative names
- AIF 1 antibody
- AIF-1 antibody
- Aif1 antibody
see all
Images
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All lanes : Anti-Iba1 antibody (ab153696) at 1/1000 dilution
Lane 1 : Mouse brain
Lane 2 : Rat brain
Lysates/proteins at 50 µg/ml per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Predicted band size: 16 kDa -
Immunoprecipitation of K562 whole cell extract (human chronic myelogenous leukemia cell line from bone marrow) using 5µg/ml ab153696 to detect Iba1 protein with secondary: anti-Rabbit IgG.
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Immunohistochemistry of paraffin-embedded rat cerebellum tissue staining Iba1 protein with ab153696 at 1/1000 dilution.
Performed with heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry of paraffin-embedded mouse cerebellum tissue staining Iba1 protein with ab153696 at 1/1000 dilution.
Performed with heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry of paraffin-embedded rat brain tissue staining Iba1 protein with ab153696 at 1/2000 dilution.
Performed with heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemical analysis of THP-1 cells (human monocytic leukemia cell line) staining cytoplasmic Iba1 protein with ab153696 in 1/1000 dilution (green). Counterstaining (blue) using fluoroshield with DAPI.
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All lanes : Anti-Iba1 antibody (ab153696) at 1/500 dilution
Lane 1 : K562 whole cell extracts
Lane 2 : THP-1 whole cell extracts
Lane 3 : HL-60 whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody at 1/10000 dilution
Developed using the ECL technique.
Predicted band size: 16 kDaHigh expression in THP-1. Low expression in K562 and HL-60.
15% SDS-PAGE.
Running condition: 80V, 15min; 140V, 40 min.
Transfer condition: Semi-dry, 18 V, 60 min (Nitrocellulose membrane).
Blocking condition: 5% non-fat milk in TBST, RT, 60 min.
Primary antibody incubation: 4°C overnight.
Washing condition: 5 ml TBST, 4 x 5 min.
ECL exposure.
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Flow cytometric analysis of THP-1 (human monocytic leukemia cell line) cell line labeling Iba1 with ab153696 at 1/50 dilution (red) compared with an unlabelled sample (black).
The sample was fixed using 4% PFA in PBS at 4℃ for 15 minutes. Cells were resuspended twice in 0.1% Triton X-100 in PBS (wash buffer), then centrifuged at 4℃ for 5 mintues. The sample was incubated with the primary antibody (1/50 in PBS) for 60 minutes at 4°C. A. Alexa Fluor® 488-conjugated anti-rabbit IgG (1/1000 in PBS) at 4oC for 30 minutes was used as the secondary antibody.
Analysed using a BD Accuri™ C6 Cytometer.
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Paraffin-embedded rat brain tissue stained for Iba1 using ab153696 at 1/500 dilution in immunohistochemical analysis. Antigen retrieval: Citrate buffer, pH 6.0, 15 min
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Flow cytometic analysis of primary murine microglia cells labeling Iba1 with ab153696 at 1.0 µg per 4×105 cells (blue), Rabbit IgG (green) , Unstained (red).
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All lanes : Anti-Iba1 antibody (ab153696) at 1/500 dilution
Lane 1 : Human Iba1 full length recombinant protein at 0.1 µg
Lane 2 : HEK293 whole cell lysate at 20 µg
Lane 3 : A431 whole cell lysate at 20 µg
Lane 4 : NIH3T3 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 whole cell lysate at 30 µg
Lane 9 : MOLT4 whole cell lysate at 30 µg
Lane 10 : THP1 whole cell lysate at 30 µg
Lane 11 : THP1 whole cell lysate, PMA treated at 30 µg
Lane 12 : Raw 264.7 whole cell lysate at 30 µg
Lane 13 : C6 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: 16 kDa
Exposure time: 1 minuteBLOCKED IN 3% MILK. For ab153696 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 ab153696 (anti-Iba1 antibody; 1/500 dilution) for 18 hours at 4°C. Antibody binding was detected using HRP-labelled anti-Rabbit IgG for 1 hour at room temperature and visualised using ECL development solution ab133406.
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Anti-Iba1 antibody (ab153696) at 1/1000 dilution + Mouse liver whole cell lysate at 50 µg
Secondary
Rabbit IgG antibody (HRP) for 1 hour at room temperature at 1/10000 dilution
Predicted band size: 16 kDa15% SDS PAGE
Running conditions: 80V for 15min then 140V for 40min
Blocking: 5% non-fat milk in TBST at room temperature for 60min.
Washing conditions: 5 ml TBST, 4 x 5min
Transfer conditions: Semi-dry, 18 V, 60min (NC membrane)
Exposure system: Trident plus Western HRP Substrate
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (27)
ab153696 has been referenced in 27 publications.
- Zhou M et al. Transcriptomic Analysis Reveals that Activating Transcription Factor 3/c-Jun/Lgals3 Axis Is Associated with Central Diabetes Insipidus after Hypothalamic Injury. Neuroendocrinology 112:874-893 (2022). PubMed: 34763342
- Zhang SS et al. Long-term running exercise improves cognitive function and promotes microglial glucose metabolism and morphological plasticity in the hippocampus of APP/PS1 mice. J Neuroinflammation 19:34 (2022). PubMed: 35123512
- Xu SF et al. Astrocyte-specific loss of lactoferrin influences neuronal structure and function by interfering with cholesterol synthesis. Glia 70:2392-2408 (2022). PubMed: 35946355
- Zhang Y et al. Deciphering the dynamic niches and regeneration-associated transcriptional program of motoneurons following peripheral nerve injury. iScience 25:104917 (2022). PubMed: 36051182
- Jin L et al. Astrocytic SARM1 promotes neuroinflammation and axonal demyelination in experimental autoimmune encephalomyelitis through inhibiting GDNF signaling. Cell Death Dis 13:759 (2022). PubMed: 36055989