WB
Western blot analysis of Iba1 on different lysates with Rabbit anti-Iba1 antibody at 1/5,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: HEK-293 cell lysate (negative), Lane 3: Mouse spleen tissue lysate, Lane 4: Rat spleen tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Iba1 with Rabbit anti-Iba1 antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Iba1 antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IF-F
pecies: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/2,000, Antigen retrieval: Not required.IF-F
Species: Rat, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/1,000, Antigen retrieval: Not required.IF-P
Species: Human, Site: Spleen, Sample: Paraffin-embedded section, Antibody concentration: 1/500.IF-P
Species: Mouse, Site: Cerebral cortex, Sample: Paraffin-embedded section, Antibody concentration: 1/500.IF-P
Species: Rat, Site: Cerebral cortex, Sample: Paraffin-embedded section, Antibody concentration: 1/500.FC
Flow cytometric analysis of THP-1 cells labeling Iba1. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Iba1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Ionized calcium-binding adapter molecule 1 (Iba1), also known as allograft inflammatory factor-1 (AIF-1), is a 147 amino acid cytoplasmic, calcium-binding protein that is thought to play a role in macrophage activation and function. Iba1, containing two EF domains, is induced by cytokines and interferons. In an unstimulated state, Iba1 colocalizes with actin, and upon stimulation, translocates to lamellipodia. It is also a marker of human microglia and is expressed by macrophages in injured skeletal muscle. The gene encoding Iba1 maps to chromosome 6p21.33 and resides in the tumor necrosis factor (TNF) cluster of genes located in the region represented by the human major histocompatibility complex (MHC). |
| Immunogen | Synthetic peptide within N-terminal human Iba1. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIF-FIF-PIHCWB |
| WB:1:5000 IHC:1:1000 ICC/IF:1:250-1:500 IF-F:1:1000-1:2000 IF-P:1:500 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| 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 |
|---|---|
| Molecular Weight(MW) | 17 kDa |
| Cellular Localization | Cytoplasm, cytoskeleton, Cell projection, ruffle membrane, Cell projection, phagocytic cup. |
WB
Western blot analysis of Iba1 on different lysates with Rabbit anti-Iba1 antibody at 1/5,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: HEK-293 cell lysate (negative), Lane 3: Mouse spleen tissue lysate, Lane 4: Rat spleen tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue with Rabbit anti-Iba1 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Iba1 with Rabbit anti-Iba1 antibody at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Iba1 antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IF-F
pecies: Mouse, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/2,000, Antigen retrieval: Not required.
IF-F
Species: Rat, Site: Cerebral cortex, Sample: Frozen section, Antibody concentration: 1/1,000, Antigen retrieval: Not required.
IF-P
Species: Human, Site: Spleen, Sample: Paraffin-embedded section, Antibody concentration: 1/500.
IF-P
Species: Mouse, Site: Cerebral cortex, Sample: Paraffin-embedded section, Antibody concentration: 1/500.
IF-P
Species: Rat, Site: Cerebral cortex, Sample: Paraffin-embedded section, Antibody concentration: 1/500.
FC
Flow cytometric analysis of THP-1 cells labeling Iba1. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:5000 IHC:1:1000 ICC/IF:1:250-1:500 IF-F:1:1000-1:2000 IF-P:1:500 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃after thawing. Aliquot store at -20℃or -80℃Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
