Recombinant Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
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 - Chimeric – BSA and Azide free
- Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free
See all Iba1 primary antibodies -
Description
Mouse monoclonal [EPR16589] to Iba1 - Chimeric – BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
The details of the immunogen for this antibody are not available.
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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.
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General notes
ab283342 is the carrier-free version of ab283319.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16589 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283342 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 |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 16 kDa.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 16 kDa. |
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 [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 kDaThis data was developed using ab283319 the same antibody clone in a different buffer formulation.
Blocking and diluting buffer: 5% NFDM/TBST
Negative control: MCF7 (HPA database)
Exposure time: 136 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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
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Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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.
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Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Chimeric – BSA and Azide free (ab283342)
This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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.
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This data was developed using ab283319 the same antibody clone in a different buffer formulation.
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
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.
Datasheets and documents
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Datasheet download
References (0)
ab283342 has not yet been referenced specifically in any publications.