Name: Anti-SNX27 antibody [1C6]
SKU: ab77799
Rating: 4 (10 reviews)

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Western blot - Anti-SNX27 antibody [1C6] (ab77799)

Key features and details

Mouse monoclonal [1C6] to SNX27
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
Reacts with: Human
Isotype: IgG1

HRP

製品名

Anti-SNX27 antibody [1C6]
SNX27 一次抗体製品一覧
製品の詳細

Mouse monoclonal [1C6] to SNX27
由来種

Mouse
アプリケーション

適用あり: Flow Cyt (Intra), WB, IP, ICC/IFmore details
種交差性

交差種: Human
交差が予測される動物種: Monkey非交差種: Mouse
免疫原

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
エピトープ

N-terminal amino acids 1-267.
ポジティブ･コントロール
- This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; HEK293; A549; MDA-MB-231. ICC/IF: Jurkat cells
特記事項

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の状態

Liquid
保存方法

Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
バッファー

pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
精製度

Protein G purified
ポリ/モノ

モノクローナル
クローン名

1C6
ミエローマ

Sp2/0
アイソタイプ

IgG1
研究分野
- Neuroscience
- Processes

Alternative Versions
- HRP Anti-SNX27 antibody [1C6] (ab203648)
Compatible Secondaries
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

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ab77799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)		Use 0.1-1µg for 10⁶ cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB	(6)	Use a concentration of 10 µg/ml. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). Abcam recommends using 3% milk as the blocking agent.
IP		Use at an assay dependent concentration.
ICC/IF	(4)	Use a concentration of 5 µg/ml.

特記事項
Flow Cyt (Intra) Use 0.1-1µg for 10⁶ cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 10 µg/ml. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa). Abcam recommends using 3% milk as the blocking agent.
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

機能

Involved in endocytic trafficking (By similarity). In T lymphocytes, participates in endocytic recycling pathway. Recruits PSCDBP and HT4R to early endosomes.
組織特異性

Expressed in cells of hematopoietic origin (at protein level).
配列類似性

Belongs to the sorting nexin family.
Contains 1 PDZ (DHR) domain.
Contains 1 PX (phox homology) domain.
Contains 1 Ras-associating domain.
ドメイン

The PDZ domain mediates the interaction with DGKZ, PSCDBP and HT4R and is responsible for vesicular localization.
細胞内局在

Cytoplasm > cytosol. Early endosome. In T-lymphocytes, recruited from the cytosol to sorting endosomes by phosphoinositide-3-kinase products.
Target information above from: UniProt accession Q96L92 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: 81609 Human
- Omim: 611541 Human
- SwissProt: Q96L92 Human
- Unigene: 192326 Human
別名
- KIAA0488 antibody
- Methamphetamine responsive transcript 1 antibody
- MGC126871 antibody
- MGC126873 antibody
- MGC20471 antibody
- MRT1 antibody
- MY014 antibody
- OTTHUMP00000042487 antibody
- RP11-98D18.12 antibody
- SNX27 antibody
- SNX27_HUMAN antibody
- Sorting nexin family member 27 antibody
- Sorting nexin-27 antibody
see all

Western blot - Anti-SNX27 antibody [1C6] (ab77799)This image is courtesy of an anonymous Abreview

Anti-SNX27 antibody [1C6] (ab77799) at 1/1000 dilution + human kidney whole cell lysate at 30 µg

Secondary
IRDye® 680RD Goat anti-Mouse IgG (H + L) at 1/15000 dilution

Performed under reducing conditions.

Predicted band size: 61 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

Exposure time: 5 minutes

4-12% bis-tris gel, MES buffer
See Abreview
Western blot - Anti-SNX27 antibody [1C6] (ab77799)

All lanes : Anti-SNX27 antibody [1C6] (ab77799) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 4 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
Lane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time: 15 minutes

Blocking: 5% Milk in TBS-T.

Two bands are observed very close together, at around 60-kDa. There are four reported isoforms of the SNX27 protein (Swissprot). We hypothesise that this antibody detects both isoform 1 and isoform 2, which are 61-kDa and 60-kDa respectively. As the immunogen is derived from the N-terminal region of SNX27, the antibody is not expected to react with isoforms 3 or 4.
Immunocytochemistry/ Immunofluorescence - Anti-SNX27 antibody [1C6] (ab77799)

ab77799 staining SNX27 in Jurkat cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab77799 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
Flow Cytometry (Intracellular) - Anti-SNX27 antibody [1C6] (ab77799)

Overlay histogram showing Jurkat cells stained with ab77799 (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 (ab77799, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunoprecipitation - Anti-SNX27 antibody [1C6] (ab77799)

SNX27 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to SNX27 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab77799.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
Band: 61kDa: SNX27.

Flow cytometry protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab77799 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。

ab77799 は 40 報の論文で使用されています。

Pei Y et al. RAB21 controls autophagy and cellular energy homeostasis by regulating retromer-mediated recycling of SLC2A1/GLUT1. Autophagy 19:1070-1086 (2023). PubMed: 35993307
Yang B et al. SNX27 suppresses SARS-CoV-2 infection by inhibiting viral lysosome/late endosome entry. Proc Natl Acad Sci U S A 119:N/A (2022). PubMed: 35022217
Simonetti B et al. SNX27-Retromer directly binds ESCPE-1 to transfer cargo proteins during endosomal recycling. PLoS Biol 20:e3001601 (2022). PubMed: 35417450
Cattin-Ortolá J et al. Sequences in the cytoplasmic tail of SARS-CoV-2 Spike facilitate expression at the cell surface and syncytia formation. Nat Commun 12:5333 (2021). PubMed: 34504087
González-Mancha N et al. Sorting Nexin 27 Enables MTOC and Secretory Machinery Translocation to the Immune Synapse. Front Immunol 12:814570 (2021). PubMed: 35095913

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 11 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (kidney)

Gel Running Conditions

Reduced Denaturing (4-12% bis-tris gel, MES buffer)

Loading amount

30 µg

Specification

kidney

Blocking step

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

Abcam user community

Verified customer

投稿 Nov 03 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (Dopaminergic Neurons)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

Dopaminergic Neurons

Blocking step

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

Abcam user community

Verified customer

投稿 Oct 19 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (U2OS)

Permeabilization

Yes - 0.5% triton-X100

Specification

U2OS

Blocking step

(agent) for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 21°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.

Abcam user community

Verified customer

投稿 Jun 17 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Permeabilization

Yes - Tx100

Specification

HeLa

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°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.

Abcam user community

Verified customer

投稿 May 19 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (HEX293T)

Gel Running Conditions

Reduced Denaturing

Loading amount

50 µg

Specification

HEX293T

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.

Abcam user community

Verified customer

投稿 May 19 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

25 µg

Specification

HeLa cells

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.

Abcam user community

Verified customer

投稿 Oct 17 2016

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 20°C

Sample

Human Cell (HeLa)

Specification

HeLa

Permeabilization

Yes - 0.1% Triton TX-100

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.

Abcam user community

Verified customer

投稿 May 23 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (HEK293T cells)

Loading amount

25 µg

Specification

HEK293T cells

Gel Running Conditions

Reduced Denaturing (4-12% SDS-PAGE gel)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Jan 24 2013

Application

Western blot

Sample

Rat Tissue lysate - whole (brain and PC12 cells)

Loading amount

40 µg

Specification

brain and PC12 cells

Gel Running Conditions

Reduced Denaturing (10%)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Mar 30 2012

Question

Hi,
Recently I have been working on SNX27 and have purchased your mouse monoclonal
antibody (ab77799, clone 1C6). This antibody is excellent, but unfortunately
I'm now doing some experiments on rat samples and this antibody
doesn't cross-react with rat SNX27. However, I notice you have another
SNX27 antibody (rabbit polyclonal; ab103354). The datasheet for this antibody
says it reacts only with human, but since the 30 amino acid immunizing antigen
differs in only 2 amino acids between the human and rat sequence, I have a
feeling it may well recognise rat.
So, I was wondering whether it may be possible to obtain a small tester
aliqout of this antibody (ab103354) to ensure it recognises rat SNX27
prior to purchasing it? I would, of course, inform you of the outcome of
this blot so you could update your website, and would happily
provide a representative blot for you to use.

Abcam community

Verified customer

Asked on Mar 12 2012

Answer

Thank you very much for your interest in our SNX27 antibodies.

To our knowledge, neither ab77799 nor ab103354 have been tested in rat yet, but that does not mean they might not work.

I absolutely agree that with an immunogen sequence similarity of 93% (i.e. 28 out of 30 amino acids) the antibody ab103354 may indeed cross-react with rat. Unfortunately, because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products. But I am happy to let you know that we have our Abcam guarantee in place (www.abcam.com/Abpromise), and for cases such as yours our testing discount programme:

As this antibody has not been tested in rat but might work, we can offer a discount off a future purchase if you buy the antibody now, test it in rat and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab103354 in rat. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering.

2. Purchase ab103354 either by phone, fax, or online (www.abcam.com).

3. Test it in rat.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews .

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab103354 turns out to be unsuitable for rat, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about these offers and I would be happy to help you further.

The Terms and Conditions can be found at: www.abcam.com/collaborationdiscount

PS: If you are happy with the results of ab77799, we would be very grateful if you could submit an Abreview about this product via the online product datasheet. You could earn quite a few Abpoints with it ;-)

Abcam Scientific Support

Answered on Mar 12 2012

1-10 of 11 Abreviews or Q&A

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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

エピトープ

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

ミエローマ

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

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

ドメイン

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

参考文献 (40)

レビューと Q&A

Western blot abreview for Anti-SNX27 antibody [1C6]

Western blot abreview for Anti-SNX27 antibody [1C6]

Immunocytochemistry/ Immunofluorescence abreview for Anti-SNX27 antibody [1C6]

Immunocytochemistry/ Immunofluorescence abreview for Anti-SNX27 antibody [1C6]

Western blot abreview for Anti-SNX27 antibody [1C6]

Western blot abreview for Anti-SNX27 antibody [1C6]

Immunocytochemistry/ Immunofluorescence

Western blot abreview for Anti-SNX27 antibody [1C6]

Western blot abreview for Anti-SNX27 antibody [1C6]