重组Anti-CD68抗体[EPR23917-164] (ab283654)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23917-164] to CD68
- Suitable for: IHC-P, IHC-Fr, ICC/IF, Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Mouse, Rat
Related conjugates and formulations
概述
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产品名称
Anti-CD68抗体[EPR23917-164]
参阅全部 CD68 一抗 -
描述
兔单克隆抗体[EPR23917-164] to CD68 -
宿主
Rabbit -
特异性
Mouse unsuitable for IHC-Fr application
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经测试应用
适用于: IHC-P, IHC-Fr, ICC/IF, Flow Cyt (Intra), WBmore details
不适用于: IP -
种属反应性
与反应: Mouse, Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Mouse spleen, RAW264.7, Mouse spleen, J774A.1, Rat liver and Rat spleen lysates. IHC-P: Mouse spleen, Mouse colon, Mouse large B lymphoma, Rat spleen, Rat lung and Rat liver tissues. IHC-Fr: and Rat liver, Rat spleen tissues. ICC/IF: J774A.1 cells. Flow Cyt: RAW 264.7 cell.
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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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23917-164 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab283654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 |
---|---|---|
IHC-P | (1) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
IHC-Fr |
1/100.
Mouse unsuitable for IHC-Fr application |
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ICC/IF |
1/50.
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Flow Cyt (Intra) |
1/500.
|
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WB |
1/1000. Predicted molecular weight: 37 kDa.
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说明 |
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IHC-Fr
1/100. Mouse unsuitable for IHC-Fr application |
ICC/IF
1/50. |
Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 37 kDa. |
靶标
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功能
Could play a role in phagocytic activities of tissue macrophages, both in intracellular lysosomal metabolism and extracellular cell-cell and cell-pathogen interactions. Binds to tissue- and organ-specific lectins or selectins, allowing homing of macrophage subsets to particular sites. Rapid recirculation of CD68 from endosomes and lysosomes to the plasma membrane may allow macrophages to crawl over selectin-bearing substrates or other cells. -
组织特异性
Highly expressed by blood monocytes and tissue macrophages. Also expressed in lymphocytes, fibroblasts and endothelial cells. Expressed in many tumor cell lines which could allow them to attach to selectins on vascular endothelium, facilitating their dissemination to secondary sites. -
序列相似性
Belongs to the LAMP family. -
翻译后修饰
N- and O-glycosylated. -
细胞定位
Cell membrane and Endosome membrane. Lysosome membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 12514 Mouse
- Entrez Gene: 287435 Rat
- SwissProt: P31996 Mouse
- Unigene: 15819 Mouse
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别名
- CD 68 antibody
- CD68 antibody
- CD68 antigen antibody
see all
图片
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All lanes : Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution
Lane 1 : Wild-type RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 :Mouse CD68 knockout RAW 264.7 cell lysate (ab280106)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 37 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
Lanes 1-2: Merged signal (red and green). Green - ab283654 observed at 100 kDa. Red - loading control ab8245 observed at 36 kDa.
ab283654 Anti-CD68 antibody [EPR23917-164] was shown to specifically react with CD68 in wild-type RAW264.7 cells. Loss of signal was observed when knockout cell line (knockout cell lysate - ab280106) was used. Wild-type and CD68 knockout samples were subjected to SDS-PAGE. ab283654 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh) tissue labeling CD68 with ab283654 at 1/100 (4.66 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on Kupffer cells of rat liver is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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All lanes : Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution
Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 37 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: mouse skeletal muscle (PMID: 28091823).
Exposure time: 37 seconds
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 cells labelling CD68 with ab283654 at 1/50 (9.32 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in J774A.1 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling CD68 with ab283654 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells in mouse colon. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh) tissue labeling CD68 with ab283654 at 1/100 (4.66 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on macrophage of rat spleen is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
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All lanes : Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution
Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 2 : RAW264.7 treated with PNGase F whole cell lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Mouse spleen treated with PNGase F tissue lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Macrosialin (CD68) is a glycoprotein and can be de-glycosylated by PNGase F. The molecular mass observed is consistent with the literature (PMID: 7680921)
Exposure time: Lane 1-2: 7.75 seconds; Lane 3-4: 48 seconds
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All lanes : Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution
Lane 1 : J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate 20
Lane 2 : Rat liver tissue lysate
Lane 3 : Rat spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Predicted band size: 37 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lane 1: 7.75 seconds; Lane 2: 125 seconds; Lane 3: 92 seconds
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Immunohistochemical analysis of paraffin-embedded Mouse large B cell lymphoma tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse large B cell lymphoma. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat spleen. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on macrophages in rat lung. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labelling CD68 with ab283654 at 1/100 (4.66 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on Kupffer cells in rat liver. The section was incubated with ab283654 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (9)
ab283654 被引用在 9 文献中.
- Ye T et al. Large extracellular vesicles secreted by human iPSC-derived MSCs ameliorate tendinopathy via regulating macrophage heterogeneity. Bioact Mater 21:194-208 (2023). PubMed: 36101856
- Rong W et al. Caspase-8 Promotes Pulmonary Hypertension by Activating Macrophage-Associated Inflammation and IL-1β (Interleukin 1β) Production. Arterioscler Thromb Vasc Biol 42:613-631 (2022). PubMed: 35387479
- Trappetti V et al. Targeted Accumulation of Macrophages Induced by Microbeam Irradiation in a Tissue-Dependent Manner. Biomedicines 10:N/A (2022). PubMed: 35453485
- Xu N et al. Adiponectin Ameliorates GMH-Induced Brain Injury by Regulating Microglia M1/M2 Polarization Via AdipoR1/APPL1/AMPK/PPARγ Signaling Pathway in Neonatal Rats. Front Immunol 13:873382 (2022). PubMed: 35720361
- Yu M et al. Metformin, Rapamycin, or Nicotinamide Mononucleotide Pretreatment Attenuate Cognitive Impairment After Cerebral Hypoperfusion by Inhibiting Microglial Phagocytosis. Front Neurol 13:903565 (2022). PubMed: 35769369
- Zhang J et al. IFN-γ enhances the efficacy of mesenchymal stromal cell-derived exosomes via miR-21 in myocardial infarction rats. Stem Cell Res Ther 13:333 (2022). PubMed: 35870960
- Liang J et al. Bioinformatics analyses of potential ACLF biological mechanisms and identification of immune-related hub genes and vital miRNAs. Sci Rep 12:14052 (2022). PubMed: 35982134
- He Y et al. HO-1 knockdown upregulates the expression of VCAM-1 to induce neutrophil recruitment during renal ischemia-reperfusion injury. Int J Mol Med 48:N/A (2021). PubMed: 34368855
- Li H et al. Rifaximin-mediated gut microbiota regulation modulates the function of microglia and protects against CUMS-induced depression-like behaviors in adolescent rat. J Neuroinflammation 18:254 (2021). PubMed: 34736493