Collagen III重组抗体[EPR17673]_Collagen III抗体(ab224532)

产品名称

Anti-Collagen III抗体[EPR17673] - BSA and Azide free
参阅全部 Collagen III 一抗
描述

兔单克隆抗体[EPR17673] to Collagen III - BSA and Azide free
宿主

Rabbit
经测试应用

适用于: Flow Cyt (Intra), WB, ICC/IF, IPmore details
种属反应性

与反应: Mouse, Rat, Human
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: Human fetal liver, fetal kidney, skin, skeletal muscle and heart lysates; HeLa, MCF7, HaCaT and A431 whole cell lysates. Mouse skeletal muscle tissue lysates, rat skeletal muscle tissue lysates, mouse heart tissue lysate, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysates. C6, RAW264.7 and PC-12 whole cell fresh lysate. ICC/IF: HeLa, MCF7, PC-12 and RAW 264.7 cells Flow Cyt (intra): HeLa, PC-12 and RAW 264.7 cells IP: HeLa, PC-12 and RAW 264.7 whole cell lysates
常规说明
ab224532 is the carrier-free version of ab184993.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.
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.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C. Do Not Freeze.
存储溶液

pH: 7.2
Constituent: PBS
无载体

是
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR17673
同种型

IgG
研究领域

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Abpromise™承诺保证使用ab224532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
Flow Cyt (Intra)		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 139 kDa).
ICC/IF		Use at an assay dependent concentration.
IP		Use at an assay dependent concentration.

说明
Flow Cyt (Intra) Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 139 kDa).
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

功能

Collagen type III occurs in most soft connective tissues along with type I collagen.
疾病相关

Defects in COL3A1 are a cause of Ehlers-Danlos syndrome type 3 (EDS3) [MIM:130020]; also known as benign hypermobility syndrome. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS3 is a form of Ehlers-Danlos syndrome characterized by marked joint hyperextensibility without skeletal deformity.
Defects in COL3A1 are the cause of Ehlers-Danlos syndrome type 4 (EDS4) [MIM:130050]. EDS is a connective tissue disorder characterized by hyperextensible skin, atrophic cutaneous scars due to tissue fragility and joint hyperlaxity. EDS4 is the most severe form of the disease. It is characterized by the joint and dermal manifestations as in other forms of the syndrome, characteristic facial features (acrogeria) in most patients, and by proneness to spontaneous rupture of bowel and large arteries. The vascular complications may affect all anatomical areas.
Defects in COL3A1 are a cause of susceptibility to aortic aneurysm abdominal (AAA) [MIM:100070]. AAA is a common multifactorial disorder characterized by permanent dilation of the abdominal aorta, usually due to degenerative changes in the aortic wall. Histologically, AAA is characterized by signs of chronic inflammation, destructive remodeling of the extracellular matrix, and depletion of vascular smooth muscle cells.
序列相似性

Belongs to the fibrillar collagen family.
Contains 1 fibrillar collagen NC1 domain.
Contains 1 VWFC domain.
翻译后修饰

Proline residues at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.
O-linked glycan consists of a Glc-Gal disaccharide bound to the oxygen atom of a post-translationally added hydroxyl group.
细胞定位

Secreted > extracellular space > extracellular matrix.
Target information above from: UniProt accession P02461 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 1281 Human
- Entrez Gene: 12825 Mouse
- Entrez Gene: 84032 Rat
- Omim: 120180 Human
- SwissProt: P02461 Human
- SwissProt: P08121 Mouse
- SwissProt: P13941 Rat
- Unigene: 443625 Human
- Unigene: 249555 Mouse
- Unigene: 3247 Rat
see all
别名
- Alpha 1 type III collagen antibody
- Alpha1 (III) collagen antibody
- CO3A1_HUMAN antibody
- COL 3A1 antibody
- COL3A1 antibody
- Collagen alpha 1(III) chain antibody
- Collagen alpha-1(III) chain antibody
- Collagen III alpha 1 chain precursor antibody
- Collagen III alpha 1 polypeptide antibody
- Collagen type III alpha 1 (Ehlers Danlos syndrome type IV autosomal dominant) antibody
- Collagen type III alpha 1 antibody
- Collagen type III alpha 1 chain antibody
- Collagen type III alpha antibody
- Collagen, fetal antibody
- EDS4A antibody
- Ehlers Danlos syndrome type IV, autosomal dominant antibody
- Fetal collagen antibody
- Type III collagen antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells labeling Collagen III with ab184993 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Western blot - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

All lanes : Anti-Collagen III antibody [EPR17673] (ab184993) at 1/1000 dilution

Lane 1 : Human fetal liver lysate at 20 µg
Lane 2 : Human fetal kidney lysate at 20 µg
Lane 3 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 5 : HaCaT (human keratinocyte cell line) whole cell lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 139 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Exposure time : Lane 1: 3 minutes; Lane 2: 15 seconds; Lane 3: 3 seconds; Lanes 4-5: 30 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID: 26017148).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunoprecipitation - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Collagen III was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184993 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184993 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: HeLa whole cell lysate 10 µg (Input).

Lane 2: ab184993 IP in HeLa whole cell lysate .

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184993 in HeLa whole cell lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5.5 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunocytochemistry/ Immunofluorescence - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Collagen III with ab184993 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunocytochemistry/ Immunofluorescence - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma small irregularly shaped cells) cell line labeling Collagen III with ab184993 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody (ab150081) at 1/1000 dilution (green).Confocal image showing cytoplasmic staining in PC-12 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody (ab150081) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunocytochemistry/ Immunofluorescence - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cell line labeling Collagen III with ab184993 at 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody (ab150081) at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, secondary is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody (ab150081) at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunoprecipitation - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Collagen III was immunoprecipitated from 0.35 mg of PC-12 (rat adrenal gland pheochromocytoma cell), whole cell lysate with ab184993 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184993 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: PC-12 whole cell lysate 10 μg (Input).

Lane 2: ab184993 IP in PC-12 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184993 in PC-12 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3.25 seconds.

Lysate was freshly made and used immediately to minimize protein degradation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Western blot - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

All lanes : Anti-Collagen III antibody [EPR17673] (ab184993) at 1/1000 dilution

Lane 1 : Human skin lysate
Lane 2 : Human skeletal muscle lysate
Lane 3 : Human heart lysate
Lane 4 : A431 (human epidermoid carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 2-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 139 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Exposure time : Lanes 1-3: 30 seconds; Lane 4: 5 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID: 26017148).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Western blot - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

All lanes : Anti-Collagen III antibody [EPR17673] (ab184993) at 1/1000 dilution

Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 5 : C6 (rat glial tumor glial cell), whole cell fresh lysate
Lane 6 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell fresh lysate
Lane 7 : PC-12 (rat adrenal gland pheochromocytoma), whole cell fresh lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 139 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Exposure time : Lanes 1-4: 92 seconds; Lanes 5-6: 48 seconds; Lane 7: 37 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

In lane 1-4, the lysates were stored at -80? prior to Western Blotting.

In lane 5-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Western blot - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

All lanes : Anti-Collagen III antibody [EPR17673] (ab184993) at 1/1000 dilution

Lane 1 : Mouse skeletal muscle tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 139 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunoprecipitation - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Collagen III was immunoprecipitated from 0.35 mg of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate with ab184993 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184993 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: RAW 264.7 whole cell lysate 10 μg (Input).

Lane 2: ab184993 IP in RAW 264.7 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184993 in RAW 264.7 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3.25 seconds.

Lysate was freshly made and used immediately to minimize protein degradation.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Flow Cytometry (Intracellular) - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Collagen III with ab184993 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Flow Cytometry (Intracellular) - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cell line labeling Collagen III with ab184993 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150081) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Flow Cytometry (Intracellular) - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling Collagen III with ab184993 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150081) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Immunoprecipitation - Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

Collagen III was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184993 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184993 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HeLa whole cell lysate 10 µg (Input).

Lane 2: ab184993 IP in HeLa whole cell lysate .

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184993 in HeLa whole cell lysate .

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184993).
Anti-Collagen III antibody [EPR17673] - BSA and Azide free (ab224532)

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

Datasheet download

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Key features and details

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概述

产品名称

描述

宿主

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

无载体

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Related Products

应用

The Abpromise guarantee

靶标

功能

疾病相关

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

Datasheet download

Certificate of Compliance

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