Anti-CD68 抗体 [EPR23917-164] (ab283654)

製品名

Anti-CD68 antibody [EPR23917-164]
CD68 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR23917-164] to CD68
由来種

Rabbit
特異性

Mouse unsuitable for IHC-Fr application
アプリケーション

適用あり: IHC-P, IHC-Fr, ICC, Flow Cyt (Intra), WBmore details
適用なし: IP
種交差性

交差種: Mouse, Rat
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- 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: J774A.1 cells. Flow Cyt: RAW 264.7 cell.
特記事項
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR23917-164
アイソタイプ

IgG
研究分野
- Neuroscience
- Development

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Isotype control
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
KO cell lines
- Mouse CD68 knockout RAW 264.7 cell line (ab280047)
KO cell lysates
- Mouse CD68 knockout RAW 264.7 cell lysate (ab280106)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab283654の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
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		1/50.
Flow Cyt (Intra)		Use at an assay dependent dilution.
WB		1/1000. Predicted molecular weight: 37 kDa.

特記事項
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 1/50.
Flow Cyt (Intra) Use at an assay dependent dilution.
WB 1/1000. Predicted molecular weight: 37 kDa.

追加情報

Is unsuitable for IP.

機能

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.
組織特異性

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.
配列類似性

Belongs to the LAMP family.
翻訳後修飾

N- and O-glycosylated.
細胞内局在

Cell membrane and Endosome membrane. Lysosome membrane.
Target information above from: UniProt accession P34810 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 12514 Mouse
- Entrez Gene: 287435 Rat
- SwissProt: P31996 Mouse
- Unigene: 15819 Mouse
別名
- CD 68 antibody
- CD68 antibody
- CD68 antigen antibody
- CD68 molecule antibody
- CD68_HUMAN antibody
- DKFZp686M18236 antibody
- gp11 antibody
- Gp110 antibody
- LAMP4 antibody
- Macrophage antigen CD68 (microsialin) antibody
- MACROPHAGE ANTIGEN CD68 antibody
- Macrosialin antibody
- SCARD1 antibody
- Scavenger receptor class D member 1 antibody
see all

Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654)

All lanes : Anti-CD68 antibody [EPR23917-164] (ab283654) at 1/1000 dilution

Lane 1 : Wild Type RAW264.7 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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.
Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunocytochemistry/ Immunofluorescence - Anti-CD68 antibody [EPR23917-164] (ab283654)

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.
Flow Cytometry (Intracellular) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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.
Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Western blot - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD68 antibody [EPR23917-164] (ab283654)

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
Anti-CD68 antibody [EPR23917-164] (ab283654)

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.

Click here to view the general protocols

SDS download

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Datasheet download

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ab283654 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab283654 は 1 報の論文で使用されています。

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

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製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

組織特異性

配列類似性

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

Certificate of Compliance

参考文献 (1)

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