Anti-RNF40 antibody [EPR18511]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (AB191309)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling RNF40 with ab191309 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of human colon is observed. Counter stained with Hematoxylin.

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

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

• IP

Supplier Data

Immunoprecipitation - Anti-RNF40 antibody [EPR18511] (AB191309)

RNF40 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab191309 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab191309 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate 10ug (Input).

Lane 2 : ab191309 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab191309 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-RNF40 antibody [EPR18511] (ab191309)

Predicted band size: 113 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (AB191309)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling RNF40 with ab191309 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of rat colon is observed. Counter stained with Hematoxylin.

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

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-RNF40 antibody [EPR18511] (AB191309)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling RNF40 with ab191309 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of mouse colon is observed. Counter stained with Hematoxylin.

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

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

• WB

Supplier Data

Western blot - Anti-RNF40 antibody [EPR18511] (AB191309)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

All lanes:

Western blot - Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney 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: 113 kDa

Observed band size: 130 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-RNF40 antibody [EPR18511] (AB191309)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1, 2 and 3 : 30 seconds; Lane 4, 5 and 6 : 15 seconds.

The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

All lanes:

Western blot - Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

mESC (Mouse embryonic stem cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 5:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 6:

HepG2 (Human liver hepatocellular carcinoma) whole 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 1/100000 dilution

Predicted band size: 113 kDa

Observed band size: 130 kDa

false

• WB

Supplier Data

Western blot - Anti-RNF40 antibody [EPR18511] (AB191309)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

All lanes:

Western blot - Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 2:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 113 kDa

Observed band size: 130 kDa

false

Exposure time: 15s