Anti-RING1 antibody [EPR13047] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.

Immunofluorescence analysis of HeLa cells labeling RING1 using ab180170 at 1/50 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling RING1 using ab180170 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling RING1 using ab180170 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.RING1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab180170 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab180170 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/10000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab180170 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab180170 in mouse cerebellum tissue lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-RING1 antibody [EPR13047] (<a href='/en-us/products/primary-antibodies/ring1-antibody-epr13047-ab180170'>ab180170</a>)

Predicted band size: 42 kDa

false

• WB

Lab

Western blot - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times :

Lanes 1-3 : 3 minutes;

Lane 4 : 1 minute.

All lanes:

Western blot - Anti-RING1 antibody [EPR13047] (<a href='/en-us/products/primary-antibodies/ring1-antibody-epr13047-ab180170'>ab180170</a>) at 1/5000 dilution

Lane 1:

Ramos (human Burkitt's lymphoma) whole cell lysate at 10 µg

Lane 2:

MOLT-4 (human acute lymphoblastic leukemia) whole cell lysate at 10 µg

Lane 3:

LNCaP (Human prostate carcinoma) whole cell lysate at 10 µg

Lane 4:

HeLa (human cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution

Predicted band size: 42 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-RING1 antibody [EPR13047] - BSA and Azide free (AB250174)

This data was developed using ab180170, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : RING1 knockout HAP1 cell lysate (20 μg)

Lane 3 : MOLT4 cell lysate (20 μg)

Lane 4 : LnCaP cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab180170 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab180170 was shown to specifically react with RING1 when RING1 knockout samples were used. Wild-type and RING1 knockout samples were subjected to SDS-PAGE. ab180170 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RING1 antibody [EPR13047] (<a href='/en-us/products/primary-antibodies/ring1-antibody-epr13047-ab180170'>ab180170</a>)

Predicted band size: 42 kDa

false