Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nuclear staining on germ cells of human testis is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

KMT1B / SUV39H2 was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab190870 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab190870 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : SH-SY5Y whole cell lysate 10μg (Input).

Lane 2 : ab190870 IP in SH-SY5Y whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab190870 in SH-SY5Y whole cell lysate.

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

Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870).

All lanes:

Immunoprecipitation - Anti-KMT1B / SUV39H2 antibody [EPR18495] (<a href='/en-us/products/primary-antibodies/kmt1b-suv39h2-antibody-epr18495-ab190870'>ab190870</a>)

Predicted band size: 47 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nuclear staining on germ cells of rat testis is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling KMT1B / SUV39H2 with ab190870 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleuarstaining on germ cells of mouse testis is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870)

Lanes 1 - 4 : Merged signal (red and green). Green - ab190870 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab190870 was shown to recognize SUV39H2 in wild-type U2-OS cells as signal was lost at the expected MW in SUV39H2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SUV39H2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab190870 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (ab240313) at 1/1000 dilution

Lane 1:

Wild-type U2-OS whole cell lysate at 20 µg

Lane 2:

SUV39H2 knockout U2-OS whole cell lysate at 20 µg

Lane 3:

HEK-293 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

• WB

Unknown

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870)

Blocking/Dilution buffer : 5% NFDM/TBST.

The full length SUV39H2 orthologues differ in size : human 410aa (UniProt Q9H5I1), mouse 477aa (UniProt Q9EQQ0) and rat 481aa (UniProt D3ZIH5).

All lanes:

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (ab240313) at 1/10000 dilution

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate 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 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa,54 kDa

true

Exposure time: 3min

• WB

Unknown

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870)

Blocking/Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (ab240313) at 1/1000 dilution

Lane 1:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 20 µg

Lane 3:

Human fetal kidney 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 1/50000 dilution

Predicted band size: 47 kDa

Observed band size: 47 kDa

true

Exposure time: 3min

• WB

Unknown

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (AB240313)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab190870)

Blocking/Dilution buffer : 5% NFDM/TBST.

The full length SUV39H2 orthologues differ in size : human 410aa (UniProt Q9H5I1), mouse 477aa (UniProt Q9EQQ0) and rat 481aa (UniProt D3ZIH5).

All lanes:

Western blot - Anti-KMT1B / SUV39H2 antibody [EPR18495] - BSA and Azide free (ab240313) at 1/5000 dilution

Lane 1:

Human testis lysate at 10 µg

Lane 2:

Mouse testis lysate at 10 µg

Lane 3:

Rat testis lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 47 kDa

true

Exposure time: 3min