Anti-Iba1 抗体 [EPR16589] - Rat IgG2a (Chimeric)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Iba1 with ab283346 at 1/5000 (0.173 μg/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 ab283346 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Iba1 with ab283346 at 1/5000 (0.173 μg/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 ab283346 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 cells labelling Iba1 with ab283346 at 1/100 (8.63 μg/ml) dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in THP-1 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control cell line : MCF7.

-ve control 1 : ab283346 at 1/100 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution.

-ve control 2 : ab179513 Anti-beta Tubulin rabbit monoclonal antibody at a 1/200 dilution followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Iba1 with ab283346 at 1/5000 (0.173 μg/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 ab283346 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

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 ab283346 at 1/10000 dilution (0.01 μg) (Red) compared with a Rat monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab)2 Anti-Rat IgG Fc (Alexa Fluor^® 488, ab150161) at 1/2000 dilution was used as the secondary antibody.

Negative control : MCF7.

• IP

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Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Iba1 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte), whole cell lysate 10 μg with ab283346 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab283346 at 1/1000 dilution. Goat Anti-Rat IgG (H+L), HRP) (ab205720) was used at 1/10000 dilution.

Lane 1 : THP-1 whole cell lysate 10 μg

Lane 2 : ab283346 IP in THP-1 whole cell lysate

Lane 3 : Rat monoclonal IgG (ab18450) instead of ab283346 in THP-1 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (ab283346)

Predicted band size: 16 kDa

Observed band size: 16 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Iba1 with ab283346 at 1/5000 (0.173 μg/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 ab283346 for 10 mins at room temperature and followed by rat IgG antibody (ab102248) 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.

• WB

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Western blot - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (AB283346)

Blocking and diluting buffer : 5% NFDM/TBST

Negative control : MCF7 (HPA database)

Exposure time : 37 seconds

All lanes:

Western blot - Anti-Iba1 antibody [EPR16589] - Microglia marker - Rat IgG2a (Chimeric) (ab283346) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

false