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離子鈣接頭蛋白單克隆抗體

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產品編號bsm-54132R
英文名稱Rabbit Anti-AIF1 (9A3)  antibody
中文名稱離子鈣接頭蛋白單克隆抗體
別    名AIF 1; AIF1; AIF1 protein; IBa1; iba-1; IBa 1; allograft inflammatory factor 1; Allograft inflammatory factor 1 splice variant G; balloon angioplasty responsive transcription; BART 1; G1; G1 putative splice variant of allograft inflamatory factor 1; IBA 1; IBA1; interferon gamma responsive transcript; Ionized calcium binding adapter molecule 1; ionized calcium-binding adapter molecule; IRT 1; IRT1; Microglia response factor; MRF1; Protein G1.  
研究領域細胞生物  免疫學  神經生物學  
抗體來源Rabbit
克隆類型Recombinant
克 隆 號9A3
交叉反應Rat,Mouse,Human
產品應用WB=1:500-1000, IHC-P=1:50-100, ICC=1:50-100, IF=1:50-100, Flow-Cyt=1:50
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
理論分子量17kDa
細胞定位細胞漿 細胞膜 
性    狀Liquid
濃    度1mg/ml
免 疫 原KLH conjugated synthetic peptide derived from human AIF1: 100-147/147 
亞    型IgG
純化方法affinity purified by Protein A
緩 沖 液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
注意事項This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMedPubMed
產品介紹Allograft Inflammatory Factor-1 (AIF1)or ionized calcium-binding adaptor molecule 1 (Iba1) is expressed selectively in microglia/macrophages and is a Ca2+-binding peptide produced by activated monocytes and microglial cells. It has been suggested that AIF1 expression is associated with chronic inflammatory processes. AIF1 is expressed by activated monocytes and might participate in a variety of pathogenic processes in the mammalian brain and in chronic transplant rejection. It has been shown to be expressed early and persistently in chronically rejecting cardiac allografts but not in cardiac syngrafts and host hearts.

Function:
Actin-binding protein that enhances membrane ruffling and RAC activation. Enhances the actin-bundling activity of LCP1. Binds calcium. Plays a role in RAC signaling and in phagocytosis. May play a role in macrophage activation and function. Promotes the proliferation of vascular smooth muscle cells and of T-lymphocytes. Enhances lymphocyte migration. Plays a role in vascular inflammation.

Subunit:
Homodimer (Potential). Monomer. Interacts with LCP1.

Subcellular Location:
Cytoplasm, cytoskeleton. Cell projection, ruffle membrane; Peripheral membrane protein; Cytoplasmic side. Note=Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.

Tissue Specificity:
Detected in T-lymphocytes and peripheral blood mononuclear cells.

Similarity:
Contains 2 EF-hand domains.

SWISS:
P55008

Gene ID:
199

產品圖片
Sample:
Lane 1: Lung (Rat) Lysate at 40 ug
Lane 2: Lung (Mouse) Lysate at 40 ug
Lane 3: Spleen (Rat) Lysate at 40 ug
Lane 4: Spleen (Mouse) Lysate at 40 ug
Lane 5: Lymph node (Rat) Lysate at 40 ug
Lane 6: Lymph node (Mouse) Lysate at 40 ug
Lane 7: Bone (Rat) Lysate at 40 ug
Lane 8: Bone (Mouse) Lysate at 40 ug
Lane 9: Cerebrum (Rat) Lysate at 40 ug
Lane 10: U937 (Human) Cell Lysate at 30 ug
Primary: Anti-AIF1 (bsm-54132R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17 kD
Observed band size: 17 kD
Sample:
Lane 1: U937 (Human) Cell Lysate at 30 ug
Lane 2: Spleen (Mouse) Lysate at 40 ug
Lane 3: Spleen (Rat) Lysate at 40 ug
Lane 4: Lymph node (Mouse) Lysate at 40 ug
Lane 5: Lymph node (Rat) Lysate at 40 ug
Lane 6: Bone (Mouse) Lysate at 40 ug
Lane 7: Bone (Rat) Lysate at 40 ug
Lane 8: Cerebrum (Mouse) Lysate at 40 ug
Lane 9: Cerebrum (Rat) Lysate at 40 ug
Primary: Anti-AIF1 (bsm-54132R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17 kD
Observed band size: 15 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:300 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human left parietal lobe); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human duodenum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
SH-SY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Iba1) monoclonal Antibody, Unconjugated (bsm-54132R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (human spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:50 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (Alexa Fluor? 488 ) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (AIF1 (9A3)) Monoclonal Antibody, Unconjugated (bsm-54132R) at 1:50 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (Alexa Fluor? 488 ) for 90 minutes, and DAPI for nuclei staining.
Blank control:THP-1.
Primary Antibody (green line): Rabbit Anti-Iba1 antibody (bsm-54132R)
Dilution: 1:50;
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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