Anti-METTL3 antibody [EPR18810] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IP

Unknown

Immunoprecipitation - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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).

All lanes:

Immunoprecipitation - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (ab221795)

Predicted band size: 64 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

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.

• WB

Supplier Data

Western blot - Anti-METTL3 antibody [EPR18810] - BSA and Azide free (AB221795)

This data was developed using ab195352, the same antibody clone in a different buffer formulation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

All lanes:

Western blot - Anti-METTL3 antibody [EPR18810] (<a href='/en-us/products/primary-antibodies/mettl3-antibody-epr18810-ab195352'>ab195352</a>) 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:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 64 kDa

Observed band size: 64 kDa

false