Name: Anti-ROCK2 + ROCK1 antibody [EP786Y]
Brand: RabMAb
SKU: ab45171
Rating: 4 (9 reviews)

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Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1
Suitable for: Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Cow, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 750 APC Carrier Free PE

製品名

Anti-ROCK2 + ROCK1 antibody [EP786Y]
ROCK2+ROCK1 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1
由来種

Rabbit
特異性

This antibody recognizes both the cleaved C-terminus of ROCK 1 (30 kDa) and full length protein (158 kDa). The immunogen used for this product shares 83% homology with ROCK2 and has been shown to bind recombinant human ROCK2, please see western blot images below.
アプリケーション

適用あり: Flow Cyt (Intra), IHC-Fr, WB, IHC-P, ICC/IF, IPmore details
種交差性

交差種: Mouse, Rat, Cow, Human
免疫原

Synthetic peptide within Human ROCK1 aa 1100-1200 (C terminal). The exact sequence is proprietary.
Database link: Q13464
ポジティブ･コントロール
- WB: Untreated, Calyculin A treated and Camptothecin treated HeLa whole cell lysate (ab150035). Jurkat, Ramos, PC-12 and RAW264.7 cell lysates. IHC-P: Human adenocarcinoma of the colon and thyroid gland carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 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

EP786Y
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
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)
Positive Controls
- Jurkat whole cell lysate (ab30128)
Recombinant Protein
- Recombinant human ROCK1 protein (ab51415)

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ab45171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
Flow Cyt (Intra)		1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr		Use at an assay dependent concentration. PubMed: 19295659
WB	(4)	1/2000 - 1/10000. Predicted molecular weight: 158 kDa. For unpurified use at 1/500.
IHC-P	(2)	1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF	(1)	1/50 - 1/500.
IP		1/40 - 1/50.

特記事項
Flow Cyt (Intra) 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration. PubMed: 19295659
WB 1/2000 - 1/10000. Predicted molecular weight: 158 kDa. For unpurified use at 1/500.
IHC-P 1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF 1/50 - 1/500.
IP 1/40 - 1/50.

細胞内局在

ROCK2: Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane. ROCK1: Cytoplasm. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole. Golgi apparatus membrane. Cell projection, bleb. Cytoplasm, cytoskeleton. Cell membrane. Cell projection, lamellipodium. Cell projection, ruffle. Associated with the mother centriole and an intercentriolar linker. Colocalizes with ITGB1BP1 and ITGB1 at the cell membrane predominantly in lamellipodia and membrane ruffles, but also in retraction fibers. Localizes at the cell membrane in an ITGB1BP1-dependent manner (By similarity). A small proportion is associated with Golgi membranes.
参照データベース
- Entrez Gene: 282041 Cow
- Entrez Gene: 785911 Cow
- Entrez Gene: 6093 Human
- Entrez Gene: 9475 Human
- Entrez Gene: 19877 Mouse
- Entrez Gene: 19878 Mouse
- Entrez Gene: 25537 Rat
- Entrez Gene: 81762 Rat
- Omim: 601702 Human
- Omim: 604002 Human
- SwissProt: Q28021 Cow
- SwissProt: Q8MIT6 Cow
- SwissProt: O75116 Human
- SwissProt: Q13464 Human
- SwissProt: P70335 Mouse
- SwissProt: P70336 Mouse
- SwissProt: Q62868 Rat
- SwissProt: Q63644 Rat
- Unigene: 306307 Human
- Unigene: 58617 Human
- Unigene: 276024 Mouse
- Unigene: 6710 Mouse
- Unigene: 88642 Rat
- Unigene: 89756 Rat
see all

Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)

Lane 1 : HeLa cell lysate - treated with Calyculin A
Lane 2 : HeLa cell lysate - treated with Camptothecin
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : Ramos cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 158 kDa
Observed band size: 158 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling ROCK2 + ROCK1 with purified ab45171 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ROCK2 + ROCK1 with purified ab45171 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150078, an Alexa Fluor^® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control: primary antibody (1/50) and secondary antibody, ab150113, an Alexa Fluor^® 488-conjugated goat anti-mouse IgG (1/500).
Flow Cytometry (Intracellular) - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

Overlay histogram showing HeLa cells stained with unpurified ab45171 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45171, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/1000 dilution

Lane 1 : Recombinant Human ROCK1 protein (aa 1114 to 1354) (30 kDa)
Lane 2 : Recombinant Human ROCK2 protein (aa 1132 to 1388) (30 kDa)

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 158 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?

Loading control: Anti-6X His tag® antibody [EPR20547] (ab213204)

Blocking buffer and concentration: 5% NFDM/TBST

Exposure Times:

Lane 1: 3 seconds

Lane 2: 7 seconds
Immunoprecipitation - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

ab45171 (purified) at 1/40 immunoprecipitating ROCK2 + ROCK1 in HeLa cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)

Lane 1 : PC-12 cell lysate
Lane 2 : RAW264.7 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 158 kDa
Observed band size: 158 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK2 + ROCK1 with unpurified ab45171.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171)

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

Country/Region

Language
Datasheet download

Download

ab45171 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。

ab45171 は 133 報の論文で使用されています。

Kang S et al. TRIM40 is a pathogenic driver of inflammatory bowel disease subverting intestinal barrier integrity. Nat Commun 14:700 (2023). PubMed: 36755029
Lin X et al. Targeting of G-protein coupled receptor 40 alleviates airway hyperresponsiveness through RhoA/ROCK1 signaling pathway in obese asthmatic mice. Respir Res 24:56 (2023). PubMed: 36803977
Sun J et al. Baicalin inhibits hepatocellular carcinoma cell growth and metastasis by suppressing ROCK1 signaling. Phytother Res 37:4117-4132 (2023). PubMed: 37246830
Liu J et al. Farnesyl diphosphate synthase exacerbates nonalcoholic steatohepatitis via the activation of AHR-CD36 axis. FASEB J 37:e23035 (2023). PubMed: 37310396
Tang F et al. RhoA/ROCK Signaling Is Involved in Pathological Retinal Neovascularization. J Vasc Res 60:183-192 (2023). PubMed: 37660689

View all Publications for this product

レビューと Q&A

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レビューを送る質問を送る

1-10 of 20 Abreviews or Q&A

Application

Western blot

Sample

Sea urchin Tissue lysate - whole (Embryo)

Gel Running Conditions

Non-reduced Denaturing (8%)

Loading amount

40 µg

Treatment

150nM Axitinib

Specification

Embryo

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Tsvia Gildor

Verified customer

投稿 Feb 21 2023

Application

Immunocytochemistry

Sample

Sea urchin Cell (Embryo)

Permeabilization

Specification

Embryo

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Tsvia Gildor

Verified customer

投稿 Jan 27 2023

Application

IHC - Wholemount

Sample

Mouse Embryo (Apical neuroepithelium)

Specification

Apical neuroepithelium

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Gabriel Galea

Verified customer

投稿 Jan 21 2020

Application

Western blot

Sample

Pig Cell lysate - whole cell (heart)

Gel Running Conditions

Reduced Denaturing

Loading amount

5 µg

Specification

heart

Blocking step

Licor as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 04 2016

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Infantile fibromatosis)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris pH 9

Permeabilization

Specification

Infantile fibromatosis

Blocking step

FBS / BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jul 04 2014

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Mouse embryo 18dpc)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris pH 9

Permeabilization

Specification

Mouse embryo 18dpc

Blocking step

FBS / BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jan 15 2014

Application

Western blot

Sample

Mouse Cell lysate - whole cell (MEF)

Gel Running Conditions

Reduced Denaturing (7.5)

Loading amount

30 µg

Treatment

TGF b1 o/n 5 ug/ul

Specification

MEF

Blocking step

(agent) for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Feb 27 2013

Question

Hi I understand, so can we try the different antibody ab143181 please.

Thanks

Abcam community

Verified customer

Asked on Sep 27 2012

Answer

Thank you for confirming these details and for your cooperation.

As requested, I have issued a free of charge replacement of alternative ab134181 Anti-ROCK1 antibody [EPR638Y]with the order number #####.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to letme know. I sincerely hope this vial will work in your experiments. If not, please do not hesitate to contact me and I suggest in this case the best way to proceed would be to provide a refund or credit note for the original antibody.

I wish you the best of luck and look forward to hearing from you with the results of the experiments with the new antibody.

Abcam Scientific Support

Answered on Sep 27 2012

Question

Hi following on from the email correspondence below, can you let me know if you have a new parent lot yet for ab45171? I really need to get the antibody working again. If not can I try an aliquot of ab134181 which appears to be a new ROCK1 antibody

Thanks

Abcam community

Verified customer

Asked on Sep 26 2012

Answer

Thank you once again for your message. Thank you also for your patience waiting for a response, I wanted to ensure I had time to review the whole case again fully before contacting you as it has been a while since we last corresponded.

I am sorry to confirm that we still only havethe same parent lot ofab45171ROCK1 antibody [EP786Y]available. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Many customers are purchasing this product and when I checked the records,we have received no other complaints in the last year for this product. It is unusual and regrettable that several vials have not worked for you.

I would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and you prefer a credit note in this case, or an alternative replacement of ab134181I will be pleased to arrange this for you.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Abcam Scientific Support

Answered on Sep 26 2012

Question

Hi we have tested the new ROCK1 antibody ab97592 using the same conditions/method we always used for ab45171 when it was working, but unfortunately the new antibody gave just non-specific staining. I was wondering if it was possible to try ab58305 as an alternative to see if this one will work?

Thanks

Abcam community

Verified customer

Asked on May 16 2012

Answer

Thank you for your message. I am sorry to hear that the alternative free of charge replacement has also not worked.

It is regrettable both the antibodies have not provided satisfactory results. I would like to reassure you that ab97592 is tested and guaranteed for IHC-P on human samples.

I am concerned that these experiments are still not working for you. In this case, I would like to investigate further Before deciding how to proceed in this case I would appreciate if you are able to confirm the following information which may help me to determine where the difficulty is:

1. Please confirm what the species and tissue typeof the sample. Are these frozen (IHC-Fr) or paraffin embedded sections (IHC-P)?

2. Please confirm how the samples are being processed. Are the samples fresh before fixation? How are they being fixed?

3. What dilution of antibody is being used? Has this been optimized?

4. How is the antibody being stored?

5. Please confirm the wash steps being used?

6. Is the current vial of secondary antibody working well with other primary antibodies? Please confirm the details for the primary antibody. Has a no primary control been included?

Many thanks for your continued cooperation. I look forward to hearing from you with the requested information.

Abcam Scientific Support

Answered on May 16 2012

1-10 of 20 Abreviews or Q&A

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Key features and details

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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

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

アプリケーション

The Abpromise guarantee

ターゲット情報

細胞内局在

参照データベース

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (133)

レビューと Q&A

Western blot abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

Immunocytochemistry abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

IHC - Wholemount abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

Western blot abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]

Western blot abreview for Anti-ROCK2 + ROCK1 antibody [EP786Y]