Name: Anti-ARPC5/p16 ARC antibody [EP1551Y]
Brand: RabMAb
SKU: ab51243
Rating: 5 (5 reviews)

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Western blot - Anti-p16 ARC antibody [EP1551Y] (ab51243)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP1551Y] to ARPC5/p16 ARC
Suitable for: ICC/IF, IP, WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

产品名称

Anti-ARPC5/p16 ARC抗体[EP1551Y]
参阅全部 ARPC5/p16 ARC 一抗
描述

兔单克隆抗体[EP1551Y] to ARPC5/p16 ARC
宿主

Rabbit
特异性

This antibody is specific to the p16 ARC protein, database link: O15511. There is no observed cross reactivity to the CDKN2A/p16INK4a protein.
经测试应用

适用于: ICC/IF, IP, WB, IHC-Pmore details
不适用于: Flow Cyt
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.
阳性对照
- WB: Human fetal, mouse and rat brain tissue lysates and HeLa, HepG2, SKBR-3 and MCF-7 cell lysates. IHC-P: Human uterus adenocarcinoma and spleen tissues. ICC/IF: Neuro-2a cells. IP: Human fetal brain tissue lysate.
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
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EP1551Y
同种型

IgG
研究领域

Alternative Versions
- Anti-ARPC5/p16 ARC antibody [EP1551Y] - BSA and Azide free (ab220800)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
Related Products
- Prestained Protein Ladder - Broad molecular weight (10 - 245 kDa) (ab116028)

The Abpromise guarantee

Abpromise™承诺保证使用ab51243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
ICC/IF		1/50 - 1/100.
IP		1/20 - 1/70.
WB	(5)	1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P		1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. We have obtained results that indicate IHC-P is unsuitable for mouse and rat species.

说明
ICC/IF 1/50 - 1/100.
IP 1/20 - 1/70.
WB 1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. We have obtained results that indicate IHC-P is unsuitable for mouse and rat species.

应用说明

Is unsuitable for Flow Cyt.

功能

Functions as component of the Arp2/3 complex which is involved in regulation of actin polymerization and together with an activating nucleation-promoting factor (NPF) mediates the formation of branched actin networks.
序列相似性

Belongs to the ARPC5 family.
细胞定位

Cytoplasm > cytoskeleton. Cell projection.
Target information above from: UniProt accession O15511 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: 10092 Human
- Entrez Gene: 67771 Mouse
- Entrez Gene: 360854 Rat
- Omim: 604227 Human
- SwissProt: O15511 Human
- SwissProt: Q9CPW4 Mouse
- SwissProt: Q4KLF8 Rat
- Unigene: 518609 Human
- Unigene: 288974 Mouse
- Unigene: 1639 Rat
see all
别名
- Actin related protein 2/3 complex subunit 5 (16 kD) antibody
- Actin related protein 2/3 complex subunit 5 antibody
- Actin related protein 2/3 complex, subunit 5 16kDa antibody
- Actin-related protein 2/3 complex subunit 5 antibody
- ARC16 antibody
- Arp2/3 complex 16 kDa subunit antibody
- Arp2/3 protein complex subunit p16 antibody
- ARPC 5 antibody
- Arpc5 antibody
- ARPC5_HUMAN antibody
- dJ127C7.3 antibody
- MGC88523 antibody
- p16 Arc antibody
- p16-ARC antibody
- RP1 127C7.3 antibody
see all

Western blot - Anti-p16 ARC antibody [EP1551Y] (ab51243)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: p16 ARC knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab51243 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab51243 was shown to specifically react with p16 ARC when p16 ARC knockout samples were used. Wild-type and p16 ARC knockout samples were subjected to SDS-PAGE. Ab51243 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p16 ARC antibody [EP1551Y] (ab51243)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling p16 ARC with purified ab51243 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-p16 ARC antibody [EP1551Y] (ab51243)

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling p16 ARC with purified ab51243 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Immunoprecipitation - Anti-p16 ARC antibody [EP1551Y] (ab51243)

ab51243 (purified) at a dilution of 1/20 immunoprecipitating p16 ARC in human fetal brain tissue lysate.

Lane 1 (input): Human fetal brain tissue lysate (10µg)

Lane 2 (+): ab51243 + human fetal brain tissue lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51243 in human fetal brain tissue lysate.

For western blotting, a HRP-conjugated anti-rabbit specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-p16 ARC antibody [EP1551Y] (ab51243)

All lanes : Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution (purified)

Lane 1 : HeLa whole cell lysate
Lane 2 : SKBR-3 whole cell lysate
Lane 3 : MCF-7 whole cell lysate
Lane 4 : HepG2 whole 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: 16 kDa
Observed band size: 16 kDa

Blocking and dilution buffer: 5% NFDM/TBST
Western blot - Anti-p16 ARC antibody [EP1551Y] (ab51243)

All lanes : Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution (purified)

Lane 1 : Human fetal brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue 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: 16 kDa
Observed band size: 16 kDa

Blocking and dilution buffer: 5% NFDM/TBST
Western blot - Anti-p16 ARC antibody [EP1551Y] (ab51243)

All lanes : Anti-ARPC5/p16 ARC antibody [EP1551Y] (ab51243) at 1/2000 dilution (unpurified)

Lane 1 : Human fetal brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 16 kDa

Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p16 ARC antibody [EP1551Y] (ab51243)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human uterus adenocarcinoma tissue labelling p16 ARC with unpurified ab51243 at a dilution of 1/50.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-p16 ARC antibody [EP1551Y] (ab51243)

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

发表研究结果有使用 ab51243？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab51243 被引用在 9 文献中.

Wang Y et al. 27-Hydroxycholesterol-Induced Dysregulation of Cholesterol Metabolism Impairs Learning and Memory Ability in ApoE ε4 Transgenic Mice. Int J Mol Sci 23:N/A (2022). PubMed: 36232940
Indra I et al. Sensing Actin Dynamics through Adherens Junctions. Cell Rep 30:2820-2833.e3 (2020). PubMed: 32101754
Wang Y et al. 27-Hydroxycholesterol Alters Synaptic Structural and Functional Plasticity in Hippocampal Neuronal Cultures. J Neuropathol Exp Neurol 78:238-247 (2019). PubMed: 30753597
Choi J et al. Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex. Cancer Sci 110:3788-3801 (2019). PubMed: 31571309
Moriya Y et al. Tumor suppressive microRNA-133a regulates novel molecular networks in lung squamous cell carcinoma. J Hum Genet 57:38-45 (2012). PubMed: 22089643
Kinoshita T et al. Actin-related protein 2/3 complex subunit 5 (ARPC5) contributes to cell migration and invasion and is directly regulated by tumor-suppressive microRNA-133a in head and neck squamous cell carcinoma. Int J Oncol 40:1770-8 (2012). PubMed: 22378351
Chang YF et al. Interaction between microRNAs and actin-associated protein Arpc5 regulates translational suppression during male germ cell differentiation. Proc Natl Acad Sci U S A 109:5750-5 (2012). ICC/IF ; Mouse . PubMed: 22447776
Hubert T et al. Actin and Arp2/3 localize at the centrosome of interphase cells. Biochem Biophys Res Commun 404:153-8 (2011). ICC/IF ; Human . PubMed: 21108927
Schmidt S et al. Kindlin-3-mediated signaling from multiple integrin classes is required for osteoclast-mediated bone resorption. J Cell Biol 192:883-97 (2011). ICC/IF ; Mouse . PubMed: 21357746

客户评价及客户问答

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1-7 of 7 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (RPE1 cell line)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

RPE1 cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · 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

提交于 Mar 10 2021

Application

Western blot

Sample

Mouse Tissue lysate - whole (Brain and Spleen)

Gel Running Conditions

Reduced Denaturing

Loading amount

50 µg

Specification

Brain and Spleen

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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.

Ester Canto Puig

Verified customer

提交于 Jul 04 2016

Application

Western blot

Loading amount

25 µg

Gel Running Conditions

Reduced Denaturing

Sample

Mouse Tissue lysate - whole (mouse lung)

Specification

mouse lung

Blocking step

Milk as blocking agent for 1 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

提交于 Feb 27 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (MCF7)

Loading amount

100000 cells

Specification

MCF7

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · 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

提交于 May 17 2012

Application

Western blot

Sample

Mouse Tissue lysate - whole (brain)

Loading amount

5 µg

Specification

brain

Gel Running Conditions

Reduced Denaturing (15% SDS-PAGE)

Blocking step

(agent) for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°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

提交于 Dec 15 2010

Question

His Question: What is the recommended protocol and in particular the antigen retrieval method for the attached p16 antibody.

Abcam community

Verified customer

Asked on Nov 01 2012

Answer

Thank you for contacing us. I have placed the protocol below, please let me know if you have any further questions.

2. Protocol
2.1. Deparaffinization/Rehydration
2.1.1. Heat slides in an oven at 65 °C for 1 hour.
2.1.2. De-paraffinize/hydrate using the following series of washes: two Xylene washes (5 min each), followed by two 100% ethanol rinses (5 min each), followed by 95% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, followed by H2O and a TBST wash for 5 min on a shaker.
2.2. Antigen Retrieval
2.2.1 Immerse slides into staining dish containing Antigen Retrieval Solution (citrate pH 6 was used)
2.2.2 Place covered staining dish into the rice cooker. Add 120 mL dH2O and press “cook”.
2.2.3 When “cook” is turned to “warm” (about 20–30 min), unplug the cooker and remove the staining dish to the bench top.
2.2.4 Allow to cool down, without cover, for 20 min.
2.3. Staining
2.3.1. Wash slides with TBST for 5 min on a shaker.
2.3.2. Inactivate endogenous peroxidase by covering tissue with 3% hydrogen peroxide for 10 min.
2.3.3. Wash slides three times with TBST (3 min each on a shaker).
2.3.4. Block slides with the blocking solution for 1 hour.
2.3.5. Dilute primary antibody in the blocking buffer per recommendation on the data sheet.
2.3.6. Apply primary antibody to each section 1/10 dilution and incubate overnight in the humidified chamber (4 ºC).
2.3.7. Wash slides three times with TBST (3 min each on a shaker).
2.3.8. Apply to each section secondary HRP-conjugated anti-rabbit antibody diluted in the blocking solution per manufacturer’s recommendation; incubate for 1 hour at room temperature.
2.3.9. Wash slides three times with TBST (3 min each on a shaker).
2.3.10. Add freshly prepared DAB substrate to the sections.
2.3.11. Incubate tissue sections with the substrate at room temperature until suitable staining develops (generally 2–5 min).
2.3.12. Rinse sections with water.
2.3.13. Counterstain with Hematoxylin.
2.3.14. Rinse sections with water.
2.3.15. Dehydrate samples using two rinses with 100% Ethanol (20 dips per rinse) followed by two rinses with Xylene (30 dips per rinse).
2.3.16. Mount coverslips on slides using Permount medium.

Abcam Scientific Support

回复于 Nov 01 2012

Question

Thank you very much for your assistance! I have placed an order for the antibody ab51243 but am now wondering if there is a control I can order, as well?

Abcam community

Verified customer

Asked on Oct 25 2012

Answer

Thank you for contacting us.

We do have control lysates for both recommended positive controls, human fetal brain tissue (ab29467) and human uterus adenocarcinoma tissue (ab44053).

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Abcam Scientific Support

回复于 Oct 25 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

特异性

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

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Related Products

应用

The Abpromise guarantee

靶标

功能

序列相似性

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (9)

客户评价及客户问答

Western blot abreview for Anti-p16 ARC antibody [EP1551Y]

Western blot abreview for Anti-p16 ARC antibody [EP1551Y]

Western blot abreview for Anti-p16 ARC antibody [EP1551Y]

Western blot abreview for Anti-p16 ARC antibody [EP1551Y]

Western blot abreview for Anti-p16 ARC antibody [EP1551Y]