Anti-METTL3 抗体 [EPR18810] - BSA and Azide free (ab221795)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18810] to METTL3 - BSA and Azide free
- Suitable for: IHC-P, IP, ICC/IF, WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-METTL3 antibody [EPR18810] - BSA and Azide free
METTL3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR18810] to METTL3 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: IHC-P, IP, ICC/IF, WB, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Pig, Non human primates -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Raji, HeLa, HEK-293, Jurkat, NCCIT, F9, Neuro-2a, LLC1, C6, RAW 264.7, PC-12 and NIH\3T3 cell lysates; human thymus lysate, mouse brain, spleen and heart lysates; rat brain lysate. IHC-P: Human bladder cancer, mouse testis and rat testis tissues. ICC/IF: HCT116 and HeLa cells. IP: HeLa cell lysate.
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特記事項
ab221795 is the carrier-free version of ab195352.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR18810 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
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ab221795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 |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 64 kDa (predicted molecular weight: 64 kDa).
Milk recommended as blocking agent. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 64 kDa (predicted molecular weight: 64 kDa). Milk recommended as blocking agent. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
N6-methyltransferase that methylates adenosine residues of some mRNAs. N6-methyladenosine (m6A), which is present at internal sites of some mRNAs, may play a role in the efficiency of mRNA splicing, transport or translation. -
組織特異性
Widely expressed at low level. Expressed in spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes. -
配列類似性
Belongs to the MT-A70-like family. -
翻訳後修飾
Phosphorylated upon DNA damage, probably by ATM or ATR. -
細胞内局在
Nucleus speckle. Colocalizes with speckles in interphase nuclei. Suggesting that it may be associated with nuclear pre-mRNA splicing components. - Information by UniProt
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参照データベース
- Entrez Gene: 56339 Human
- Entrez Gene: 56335 Mouse
- Entrez Gene: 361035 Rat
- Omim: 612472 Human
- SwissProt: Q86U44 Human
- SwissProt: Q8C3P7 Mouse
- Unigene: 168799 Human
- Unigene: 271759 Mouse
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別名
- adoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase antibody
- IME4 antibody
- M6A antibody
see all
画像
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All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution
Lane 1 : Mouse brain lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 2 : Mouse brain lysate prepared in RIPA lysis method at 20 mg/ml
Lane 3 : Mouse heart lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 4 : Mouse heart lysate prepared in RIPA lysis method at 20 mg/ml
Lane 5 : Mouse kidney lysate prepared in RIPA lysis method at 20 µg
Lanes 6-7 : Mouse spleen lysate prepared in spleen lysis method at 20 µg
Lane 8 : Rat brain lysate prepared in RIPA lysis method at 20 µg
Lane 9 : Rat kidney lysate prepared in RIPA lysis method at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunohistochemical analysis of paraffin-embedded Rat skin tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Rat skin. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Inset image: negative control - secondary antibody only. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Rat testis. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Inset image: negative control - secondary antibody only. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunohistochemical analysis of paraffin-embedded Mouse skin labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Mouse skin. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Inset image: negative control - secondary antibody only. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on Mouse testis. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Inset image: negative control - secondary antibody only. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling METTL3 using ab195352 at 1/2000 dilution, followed by a Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstain: Hematoxylin. Nuclear staining on human ovarian cancer. The section was incubated with ab195352 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Perform Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Inset image: negative control - secondary antibody only. -
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling METTL3 with ab195352 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NIH/3T3 cells. The nuclear counter stain is DAPI (blue). Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) ab195889 was used at 1/200 dilution as the counterstain.
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ab195352 staining METTL3 in the human cell line HEK-293 (Human epithelial cell line from embryonic kidney) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a 1/70 dilution, followed by Goat-Anti Rabbit IgG (Alexa Fluor® 488) secondary antibody at a 1/2000 dilution.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
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Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on human bladder cancer was observed.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on mouse testis was observed.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling METTL3 using ab195352 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Counterstain: Hematoxylin.
Inset image: negative control obtained using PBS instead of ab195352, and secondary antibody only.
Note: Nuclear staining on rat testis was observed.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling METTL3 with ab195352 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
1. ab195352 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
2. Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by 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 (ab195352).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT116 (Human colorectal carcinoma cell line) cells labeling METTL3 with ab195352 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HCT116 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
1. ab195352 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (AlexaFluor®594) (ab150120) secondary antibody at 1/1000 dilution.
2. anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by 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 (ab195352).
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METTL3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab195352 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab195352 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Input: 10µg of HeLa whole cell lysate.
Lane 2: HeLa whole cell lysate following IP with ab195352.
Lane 3: Negative control: IP using Rabbit monoclonal IgG (ab172730) instead of ab195352 in HeLa whole cell lysates.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. 1 second exposure.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab195352).
プロトコール
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (2)
ab221795 は 2 報の論文で使用されています。
- Xue P et al. Increased METTL3-mediated m6A methylation inhibits embryo implantation by repressing HOXA10 expression in recurrent implantation failure. Reprod Biol Endocrinol 19:187 (2021). PubMed: 34906165
- Chang G et al. YTHDF3 Induces the Translation of m6A-Enriched Gene Transcripts to Promote Breast Cancer Brain Metastasis. Cancer Cell 38:857-871.e7 (2020). PubMed: 33125861