Antibodies, Proteins, Kits and Reagents for Life Science

365 193 ₸ Product size

100 µl 365 193 ₸ 10 µl 134 016 ₸

Order now and get it on Tuesday March 02, 2021

Anti-METTL3 antibody [EPR18810] (ab195352)

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
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-METTL3 antibody [EPR18810]
See all METTL3 primary antibodies
Description

Rabbit monoclonal [EPR18810] to METTL3
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- 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.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18810
Isotype

IgG
Research areas

Western blot - Anti-METTL3 antibody [EPR18810] (ab195352) This image is courtesy of an Abreview by Pedro Batista.

All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution

Lane 1 : Mouse embryonic stem cells
Lane 2 : Mouse embryonic stem cells (METTL3 Knock-out)

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat anti-Rabbit Polyclonal at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 64 kDa

Exposure time: 5 minutes

Blocking step: 5% Milk for 30 minutes at 25^oC.
See Abreview
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

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
Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry - Anti-METTL3 antibody [EPR18810] (ab195352)

ab195352 staining METTL3 in the human cell line HEK-293 (Human epithelial cell line from embryonic kidney) by 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)
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/5000 dilution

Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 3 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 5 : NCCIT (human pluripotent embryonal carcinoma) whole cell lysate
Lane 6 : F9 (Mouse embyro testicular cancer cell line) whole cell lysate
Lane 7 : Neuro-2a (Mouse neuroblastoma cells) whole cell lysate
Lane 8 : LLC1 (Mouse lung carcinoma) 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: 64 kDa
Observed band size: 64 kDa

Exposure time: 10 seconds

5% NFDM/TBST: Blocking and diluting buffer.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

All lanes : Anti-METTL3 antibody [EPR18810] (ab195352) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 64 kDa
Observed band size: 64 kDa

Exposure time: 2 seconds

5% NFDM/TBST: Blocking and diluting buffer.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] (ab195352)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-METTL3 antibody [EPR18810] (ab195352)

Anti-METTL3 antibody [EPR18810] (ab195352) at 1/5000 dilution + Human thymus lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 64 kDa
Observed band size: 64 kDa

Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.
Immunoprecipitation - Anti-METTL3 antibody [EPR18810] (ab195352)

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.
Anti-METTL3 antibody [EPR18810] (ab195352)

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

Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-METTL3 antibody [EPR18810] (ab195352)

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Images

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