Anti-RILPL1 抗体 [MJF-R41-21] (BSA and Azide free) (ab302493)

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Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [MJF-R41-21] to RILPL1 - BSA and Azide free
Suitable for: ICC/IF, WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Unconjugated

製品名

Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free)
RILPL1 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [MJF-R41-21] to RILPL1 - BSA and Azide free
由来種

Rabbit
アプリケーション

適用あり: ICC/IF, WB, IPmore details
適用なし: Flow Cyt,IHC-Fr or IHC-P
種交差性

交差種: Mouse, Rat, Human
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: Wild-type A549 whole cell lysate. HeLa, NIH/3T3, C6, SH-SY5Y, HEK-293, Neuro-2a, and PC-12 whole cell lysates; rat and mouse brain tissue lysate. Human cerebellum, hypothalamus and heart tissue lysate. ICC/IF: SH-SY5Y and Neuro-2a cells. IP: A549 whole cell lysate, mouse and rat brain tissue lysates.
特記事項
ab302493 is the carrier-free version of ab302492.

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.
This antibody was developed with support from The Michael J. Fox Foundation.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C.
バッファー

pH: 7.20
Constituent: 100% PBS
キャリア・フリー

はい
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

MJF-R41-21
アイソタイプ

IgG
研究分野

Alternative Versions
Compatible Secondaries
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保証は、次のテスト済みアプリケーションにおけるab302493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
ICC/IF		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration. Detects a band of approximately 47, 42 kDa (predicted molecular weight: 47 kDa).
IP		Use at an assay dependent concentration.

特記事項
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 47, 42 kDa (predicted molecular weight: 47 kDa).
IP Use at an assay dependent concentration.

追加情報

Is unsuitable for Flow Cyt,IHC-Fr or IHC-P.

機能

Neuroprotective protein, which acts by sequestring GAPDH in the cytosol and prevent the apoptotic function of GAPDH in the nucleus. Competes with SIAH1 for binding GAPDH (By similarity). Does not regulate lysosomal morphology and distribution.
組織特異性

Widely expressed. Expressed at lower level in liver and kidney.
配列類似性

Belongs to the RILPL family.
Contains 1 RILP-like domain.
翻訳後修飾

S-nitrosylation is required for the interaction with GAPDH.
細胞内局在

Cytoplasm > cytosol.
Target information above from: UniProt accession Q5EBL4 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: 353116 Human
- Entrez Gene: 75695 Mouse
- Entrez Gene: 304469 Rat
- GenBank: NM_178314.3 Human
- Omim: 614092 Human
- SwissProt: Q5EBL4 Human
- SwissProt: Q9JJC6 Mouse
- SwissProt: D3ZUQ0 Rat
- Unigene: 530315 Human
- Unigene: 41180 Mouse
- Unigene: 62627 Rat
see all
別名
- GOSPEL antibody
- MGC105128 antibody
- MGC99793 antibody
- Rab interacting lysosomal protein like 1 antibody
- Rab-interacting lysosomal-like protein 1 antibody
- RILP like protein 1 antibody
- RILP-like protein 1 antibody
- RILPL 1 antibody
- rilpl1 antibody
- RIPL1_HUMAN antibody
- RLP1 antibody
see all

Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution

Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 2 : RILPL1 knockout A549 whole cell lysate
Lane 3 : HeLa (human cervical adenocarcinoma epithelial cell), whole cell lysate

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

Performed under reducing conditions.

Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer: Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

False colour image of Western blot: Anti-RILPL1 antibody [MJF-R41-21] (ab302492) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab302492 was shown to bind specifically to RILPL1. Two bands were observed at 47/42 kDa in wild-type A549 cell lysates whereas no signal observed at this size in RILPL1 knockout cell line. To generate this image, wild-type and RILPL1 knockout A549 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel and then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

The doublet bands represent two isoforms of RILPL1.

Wild-type and RILPL1 knockout A549 cell lysates were kindly provided by Dr. Dario Alessi, University of Dundee.
Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution

Lane 1 : Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 2 : RILPL1 knockout A549 whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat kidney tissue lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer: 5% NFDM/TBST.

The doublet bands represent two isoforms of RILPL1.

Negative control: kidney (PMID: 14668488)

Wild-type and RILPL1 knockout A549 cell lysates were kindly provided by Dr. Dario Alessi, University of Dundee.
Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 47 kDa
Observed band size: 47 kDa

Exposure time: 103 seconds

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer: 5% NFDM/TBST.

Negative control: kidney, liver (PMID: 14668488).
Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate
Lane 2 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?

Exposure time: 37 seconds

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer: 5% NFDM/TBST.

The doublet bands represent two isoforms of RILPL1.
Low expression: U937 (Human Protein Atlas).
Western blot - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

All lanes : Anti-RILPL1 antibody [MJF-R41-21] (ab302492) at 1/1000 dilution

Lane 1 : Human cerebellum tissue lysate
Lane 2 : Human hypothalamus tissue lysate
Lane 3 : Human heart tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 47 kDa
Observed band size: 42,47 kDa why is the actual band size different from the predicted?

Exposure time: 26 seconds

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Blocking / Diluting buffer: 5% NFDM/TBST.

The doublet bands represent two isoforms of RILPL1.
Immunocytochemistry/ Immunofluorescence - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (ab302493)

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) labelling RILPL1 with ab302492 at 1/250 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing centrosome and cytoplasmic staining in SH-SY5Y cell line. The nuclear counterstain is DAPI (blue).

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

Low expression: U-937
Immunocytochemistry/ Immunofluorescence - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (ab302493)

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) labelling RILPL1 with ab302492 at 1/250 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (green). Confocal image showing centrosome staining in Neuro-2a cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (red) at 1/200 dilution.
Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

RILPL1 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell), whole cell lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 (Input): A549 whole cell lysate 10 μg

Lane 2 (+): A549 whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in A549 whole cell lysate

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

RILPL1 was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 (Input): Mouse brain tissue lysate 10 μg

Lane 2 (+): Mouse brain tissue lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in mouse brain tissue lysate

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

Exposure time: 3 minutes.
Immunoprecipitation - Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

This data was developed using ab302492, the same antibody clone in a different buffer formulation.

RILPL1 was immunoprecipitated from 0.35 mg rat brain tissue lysate with ab302492 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302492 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 (Input): Rat brain tissue lysate 10 μg

Lane 2 (+): Rat brain tissue lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab302492 in rat brain tissue lysate

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Anti-RILPL1 antibody [MJF-R41-21] (BSA and Azide free) (AB302493)

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

Download

ab302493 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。

ab302493 は論文での使用が確認できていません。

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

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

キャリア・フリー

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

Isotype control

Related Products

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

Datasheet download

Certificate of Compliance

参考文献 (0)

レビューと Q&A