Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (Chimeric) - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (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.

• IP

Supplier Data

Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (Chimeric) - BSA and Azide free (AB283342)

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

All lanes:

Immunoprecipitation - Anti-Iba1 antibody [EPR16589] - Microglia marker - Mouse IgG1 (Chimeric) (<a href='/en-us/products/primary-antibodies/iba1-antibody-epr16589-microglia-marker-mouse-igg1-chimeric-ab283319'>ab283319</a>)

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] - Mouse IgG1 (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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (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

• WB

Lab

Western blot - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (Chimeric) - BSA and Azide free (AB283342)

This 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

All lanes:

Western blot - Anti-Iba1 antibody [EPR16589] - Mouse IgG1 (Chimeric) - BSA and Azide free (ab283342) 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:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 16 kDa

false