重组Anti-Iba1抗体[EPR16588] (ab178846)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16588] to Iba1
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Iba1抗体[EPR16588]
参阅全部 Iba1 一抗 -
描述
兔单克隆抗体[EPR16588] to Iba1 -
宿主
Rabbit -
特异性
For ab178846 Abcam recommends blocking in 3% milk for cleanest results in WB. Blocking with BSA gives slightly higher background. -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HL-60, THP-1, U937, RAW 264.7 and NR8383 whole cell lysates; Human, mouse and rat spleen lysates; Mouse testis and liver lysates; Rat and mouse hippocampus and brain tissue lysates. IHC-P: Human Cerebral cortex, human hippocampus; Rat and mouse normal brain tissues. Flow Cyt (intra): U937 cells, Raw264.7. IHC-Fr: Mouse Cerebral cortex tissue. ICC/IF: Rat hippocampal mixed glia.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR16588 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-Iba1 antibody [EPR16588] - BSA and Azide free (ab220815)
- Alexa Fluor® 488 Anti-Iba1 antibody [EPR16588] (ab225260)
- Alexa Fluor® 647 Anti-Iba1 antibody [EPR16588] (ab225261)
- Anti-Iba1 antibody [EPR16588] - Goat IgG (Chimeric) (ab289874)
- Anti-Iba1 antibody [EPR16588] – Rat IgG2a (Chimeric) (ab300156)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab178846于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/160.
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IHC-P | (18) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (2) |
1/500 - 1/2000. Detects a band of approximately 10, 15 kDa (predicted molecular weight: 17 kDa).
Abcam recommends blocking in 3% milk for cleanest results in WB. Blocking with BSA gives slightly higher background. |
ICC/IF | (6) |
Use a concentration of 0.1 - 1 µg/ml.
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说明 |
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Flow Cyt (Intra)
1/160. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/500 - 1/2000. Detects a band of approximately 10, 15 kDa (predicted molecular weight: 17 kDa). Abcam recommends blocking in 3% milk for cleanest results in WB. Blocking with BSA gives slightly higher background. |
ICC/IF
Use a concentration of 0.1 - 1 µg/ml. |
靶标
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功能
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. -
组织特异性
Detected in T-lymphocytes and peripheral blood mononuclear cells. -
序列相似性
Contains 2 EF-hand domains. -
翻译后修饰
Phosphorylated on serine residues. -
细胞定位
Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis. - Information by UniProt
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数据库链接
- Entrez Gene: 199 Human
- Entrez Gene: 11629 Mouse
- Entrez Gene: 29427 Rat
- Omim: 601833 Human
- SwissProt: P55008 Human
- SwissProt: O70200 Mouse
- SwissProt: P55009 Rat
- Unigene: 76364 Human
see all -
别名
- AIF 1 antibody
- AIF-1 antibody
- Aif1 antibody
see all
图片
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Flow cytometry overlay histogram showing left Raw264.7 positive cells and right negative NIH3T3 stained with ab178846 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab178846) (1x 106 in 100μl at 0.2μg/ml (1/9850)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-Iba1 antibody [EPR16588] (ab178846) at 1/1000 dilution
Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse hippocampus tissue lysate
Lane 4 : Rat spleen tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat hippocampus tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDa
Exposure time: 40 secondsBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
IBA1 is a relatively minor protein of brain and is much more abundant in spleen (PMID: 8912632, PMID: 29232670). We suggest loading higher amount of brain lysate or using lower dilution of antibody for detecting signal in brain related lysates.
ab181602 was used as loading control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] (ab178846)
IHC image of Iba1 staining in rat normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] (ab178846)
IHC image of Iba1 staining in human normal hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-Iba1 antibody [EPR16588] (ab178846)
Lane 1 : Mouse testis
Lane 2 : Mouse liver
Lane 3 : Rat testis
Lane 4 : Rat liver
Predicted band size: 17 kDa -
Intracellular Flow Cytometry analysis of2% paraformaldehyde fixed U937 (human histiocytic lymphoma cell line) cells labeling Iba1with ab178846 at 1/160 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).
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Different batches of ab178846 were tested on THP-1 (Human monocytic leukemia monocyte) lysate at 0.02 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 15 kDa.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] (ab178846)
IHC image of Iba1 staining in mouse normal brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab178846, 1/2000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-Iba1 antibody [EPR16588] (ab178846) at 1/500 dilution
Lane 1 : Human Iba1 recombinant protein at 0.1 µg
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 3 : A431 (human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) 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 (human histiocytic lymphoma cell line) whole cell lysate at 30 µg
Lane 9 : MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate at 20 µg
Lane 10 : THP-1 (human monocytic leukemia cell line) whole cell lysate at 30 µg
Lane 11 : THP-1 whole cell lysate, PMA treated at 30 µg
Lane 12 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 30 µg
Lane 13 : C6 (rat glial tumor cell line) whole cell lysate at 30 µg
Lane 14 : NR8383 whole cell lysate at 30 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 17 kDa
Exposure time: 1 minuteAbcam 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 ab178846 (anti-Iba1 antibody; 1/500 dilution) for 18 hours at 4°C. Antibody binding was detected using ab97040 (HRP-labelled goat anti-mouse IgG) at 1:50,000 dilution for 1 hour at room temperature and visualised using ECL development solution ab133406.
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Immunofluorescence staining of Iba-1 using ab178846 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton-X-100 (in PBS) for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab178846 at 0.1 μg/ml and ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown. The antibody ab178846 gave comparable results using MeOH fixation (100%, 5 min).
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0.1% Triton-X 100 permeabilized paraformaldehyde-fixed Mouse cell Microglia cells labeling Iba1 (green) using ab178846 at 1/500 dilution in ICC/IF analysis.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] (ab178846)This image was courtesy of an annonymous Abreview
Formaldehyde-fixed, paraffin-embedded cynomolgus monkey brain tissue stained for Iba1 using ab178846 at 1/6000 dilution in immunohistochemical analysis.
Antigen Retrieval: Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
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All lanes : Anti-Iba1 antibody [EPR16588] (ab178846) at 1/10000 dilution
Lane 1 : THP-1 (human monocytic leukemia cell line) whole cell lysate
Lane 2 : U937 (human histiocytic lymphoma cell line) whole cell lysate
Lane 3 : Human spleen whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 17 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM /TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Iba1 antibody [EPR16588] (ab178846)
Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Iba1 with ab178846 at a 1/2000 dilution showing cytoplasm and nuclear staining on Glial cells. Counter stained with hematoxylin. Prediluted HRP Polymer for Rabbit/Mouse IgG was used as the secondary aantibody. Negative control also shown.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Anti-Iba1 antibody [EPR16588] (ab178846) at 1/2000 dilution + HL-60 (human promyelocytic leukemia cell line) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 17 kDa
Observed band size: 10, 15 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration:
5% NFDM /TBST.
Based on sequence analysis, ab178846 recognizes 2 isoforms with the predicted MWs of 17KDa and 11KDa, respectively.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (285)
ab178846 被引用在 285 文献中.
- Saponara E et al. Loss of Hepatic Leucine-Rich Repeat-Containing G-Protein Coupled Receptors 4 and 5 Promotes Nonalcoholic Fatty Liver Disease. Am J Pathol 193:161-181 (2023). PubMed: 36410420
- Bellut M et al. Delayed NLRP3 inflammasome inhibition ameliorates subacute stroke progression in mice. J Neuroinflammation 20:4 (2023). PubMed: 36600259
- Liang Z et al. Long-Term High-Fat Diet Consumption Induces Cognitive Decline Accompanied by Tau Hyper-Phosphorylation and Microglial Activation in Aging. Nutrients 15:N/A (2023). PubMed: 36615907
- Smith DC et al. Deletion of the Alzheimer's disease risk gene Abi3 locus results in obesity and systemic metabolic disruption in mice. Front Aging Neurosci 14:1035572 (2022). PubMed: 36620768
- Yang F et al. Glucagon-like Peptide 1 Receptor Activation Inhibits Microglial Pyroptosis via Promoting Mitophagy to Alleviate Depression-like Behaviors in Diabetic Mice. Nutrients 15:N/A (2022). PubMed: 36615696