Anti-Iba1 antibody - Microglia marker

• Flow Cyt

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Flow Cytometry - Anti-Iba1 antibody - Microglia marker (AB153696)

Flow cytometry analysis of THP-1 cells detecting Iba1 using ab153696 at a 1/50 dilution (Red). Unlabelled sample was used as a control (Black).
Acquisition of 20,000 events were collected for FACS analysis.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Iba1 antibody - Microglia marker (AB153696)

Immunofluorescence analysis of THP-1 cells fixed in 4% paraformaldehyde at RT for 15 min labelling Iba1 protein at cell periphery using ab153696 at a 1/500 dilution (Green). DAPI was used for nuclear staining.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

Immunohistochemical analysis of paraffin-embedded rat brain tissue stained for Iba1 protein at cell membrane and cytoplasm with ab153696 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

Immunohistochemical analysis of paraffin-embedded mouse brain tissue stained for Iba1 protein at cell membrane and cytoplasm with ab153696 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

Immunohistochemical analysis of paraffin-embedded mouse brain tissue stained for Iba1 protein at cell membrane and cytoplasm with ab153696 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody - Microglia marker (AB153696)

Immunohistochemical analysis of paraffin-embedded rat brain tissue stained for Iba1 protein at cell membrane and cytoplasm with ab153696 at a 1/500 dilution.

Antigen Retrieval : Citrate buffer, pH 6.0, 15 min.

• WB

Lab

Western blot - Anti-Iba1 antibody - Microglia marker (AB153696)

BLOCKED 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.

All lanes:

Western blot - Anti-Iba1 antibody - Microglia marker (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

Predicted band size: 16 kDa

true

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-Iba1 antibody - Microglia marker (AB153696)

Samples were separated by 15% SDS-PAGE. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.

All lanes:

Western blot - Anti-Iba1 antibody - Microglia marker (ab153696) at 1/5000 dilution

Lane 1:

K562 whole cell extracts at 30 µg

Lane 2:

THP-1 whole cell extracts at 30 µg

Lane 3:

HL-60 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Observed band size: 17 kDa

false

• WB

Supplier Data

Western blot - Anti-Iba1 antibody - Microglia marker (AB153696)

Samples were separated by 15% SDS-PAGE.

All lanes:

Western blot - Anti-Iba1 antibody - Microglia marker (ab153696) at 1/500 dilution

Lane 1:

Mouse brain tissue extracts at 50 µg

Lane 2:

Rat brain tissue extracts at 50 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Observed band size: 17 kDa

false

• Flow Cyt

Unknown

Flow Cytometry - Anti-Iba1 antibody - Microglia marker (AB153696)

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).

• IP

Supplier Data

Immunoprecipitation - Anti-Iba1 antibody - Microglia marker (AB153696)

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.

All lanes:

Immunoprecipitation - Anti-Iba1 antibody - Microglia marker (ab153696)

Predicted band size: 16 kDa

false

• WB

CiteAb

Western blot - Anti-Iba1 antibody - Microglia marker (AB153696)

Iba1 western blot using anti-Iba1 antibody ab153696. Publication image and figure legend from Fu, H., Zhao, Y., et al., 2020, Cell Death Dis, PubMed 32661227.

ab153696 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab153696 please see the product overview.

CSF1R inhibition reduces microglial accumulation induced by SCI.a Schematics of experimental design showing the timeline of microglia depletion, spinal cord crush injury, and sacrifice. b Western blot analysis of spinal cord homogenates from sham-operated, injured with control or PLX3397-treated mice (n = 4 per group) for steady state levels of the microglia marker Iba1. c Quantification of b showing a robust increase in levels of Iba1 expression at 7 days following SCI. CSF1R inhibition reverses the increase induced by SCI. d–f Representative immunofluorescence images of mouse spinal cord sections showing Iba1+ microglia. The changes in number of Iba1+ microglia consist with the changes of Iba1 expression levels. Data are expressed as mean ± SD. Scale bars : e, f in f 100 µm.

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