PRODUCT CODE: ET1705-78

Recombinant Iba1 Monoclonal Antibody (ET1705-78)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Iba1 on THP-1 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Iba1 on THP-1 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of Iba1 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-78, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat lung tissue using anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue using anti-Iba1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-78, 1/50) for 30 minutes 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.
  • Flow cytometric analysis of Iba1 was done on THP-1 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-78, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Iba1 on THP-1 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1705-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

  • IP

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Iba1 Monoclonal Antibody (ET1705-78)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

THP-1 cell lysates, SH-SY5Y, rat lung tissue, human lung carcinoma tissue, human spleen tissue, mouse brain tissue, mouse spleen tissue, THP-1.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JM36-62

PROPERTIES

Form

Liquid

Storage Condition

Store at +4Cafter thawing. Aliquot store at -20Cor -80CAvoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

17 kDa

Isotype

IgG

APPLICATION DILUTION

  • ICC/IF

  • 1:50-1:100

  • IHC-P

  • 1:100-1:500

  • FC

  • 1:50-1:200

  • IP

  • 1:10-1:50 WB

TARGET

UNIPROT #

PROTEIN NAME

Iba1

SYNONYMS

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

TISSUE SPECIFICITY

Detected in T-lymphocytes and peripheral blood mononuclear cells.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated on serine residues.

SUBCELLULAR LOCATION

Ruffle membrane, cytoskeleton, phagocytic cup.

FUNCTION

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).