IBA1 Recombinant antibody

IBA1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF-P, IF-Fro, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF-P, IF-Fro, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

4D5

Cat no : 81728-1-RR

Synonyms

AIF1, G1, allograft inflammatory factor 1, AIF-1, AIF 1

---

Validation Data Gallery

• 

  WB analysis of THP-1 using 81728-1-RR

  THP-1 cells were subjected to SDS PAGE followed by western blot with 81728-1-RR (IBA1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 81728-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81728-1-RR (IBA1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of rat brain using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse liver using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse liver using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded human liver tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 81728-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 81728-1-RR (IBA1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of rat brain using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using IBA1 antibody (81728-1-RR, Clone: 4D5 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of rat brain using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed rat brain tissue using IBA1 antibody (81728-1-RR, Clone: 4D5 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  IF Staining of rat brain using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded rat brain tissue using IBA1 antibody (81728-1-RR, Clone: 4D5 ) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of rat brain using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded rat brain tissue using IBA1 antibody (81728-1-RR, Clone: 4D5 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 750-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR006). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of adult mouse cerebellum tissue using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed adult mouse cerebellum tissue using IBA1 antibody (81728-1-RR) at dilution of 1:500 and Alexa Fluor 594 AffiniPure Donkey Anti-Rabbit IgG (H+L). Heat mediated antigen retrieval with PBS buffer (pH 7.4). (Image produced by Watt Lab at McGill University)

• 

  IF Staining of mouse brain using 81728-1-RR

  Immunofluorescent analysis of (4% PFA) fixed frozen OCT-embedded mouse brain tissue using IBA1 antibody (81728-1-RR, Clone: 4D5 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

• 

  FC experiment of THP-1 using 81728-1-RR

  1x10^6 THP-1 cells were intracellularly stained with 0.4 ug IBA1 Recombinant antibody (81728-1-RR, Clone:4D5) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

---

Tested Applications

• Positive WB detected in: THP-1 cells, U-937 cells, HL-60 cells
• Positive IHC detected in: rat brain tissue, human liver tissue, mouse brain tissue, mouse liver tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: rat brain tissue
• Positive IF-Fro detected in: adult mouse cerebellum tissue, mouse brain tissue
• Positive FC (Intra) detected in: THP-1 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)-P: IF-P : 1:250-1:1000
• Immunofluorescence (IF)-FRO: IF-FRO : 1:250-1:1000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

81728-1-RR targets IBA1 in WB, IHC, IF-P, IF-Fro, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: IBA1 fusion protein Ag1363
• Full Name: allograft inflammatory factor 1
• Calculated Molecular Weight: 17 kDa
• Observed Molecular Weight: 17 kDa
• GenBank Accession Number: BC009474
• Gene Symbol: IBA1
• Gene ID (NCBI): 199
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

IBA1 is a 143 amino acid cytoplasmic, inflammation response scaffold protein. It is constitutively expressed in monocytes and macrophages and is known to be involved in macrophage activation. It is a marker of activated macrophage. Expression of IBA1 is up-regulated in activated microglia following facial nerve axotomy, ischemia, and several brain diseases, thereby implicating it in the activated phenotypes of microglia.