Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] - BSA and Azide free (AB251485)

This data was developed using ab208026, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 μM Ionomycin for 2 hours, labeling hnRNP A1 (citrulline R92) with ab208026 at 1/600 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 647) (150079) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] - BSA and Azide free (AB251485)

This data was developed using ab208026, the same antibody clone in a different buffer formulation.

hnRNP A1 (citrulline R92) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 μM Ionomycin for 2 hours, whole cell lysate with ab208026 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208026 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HEK-293T transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 μM Ionomycin for 2 hours, whole cell lysate 10 μg (Input).

Lane 2 : ab208026 IP in HEK-293T transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 μM Ionomycin for 2 hours, whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab208026 in HEK-293T transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 μM Ionomycin for 2 hours, whole cell lysate.

Exposure time :Less than 1 second.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] (<a href='/en-us/products/primary-antibodies/hnrnp-a1-citrulline-r92-antibody-epr20174-ab208026'>ab208026</a>)

Predicted band size: 38 kDa

false

• WB

Supplier Data

Western blot - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] - BSA and Azide free (AB251485)

This data was developed using ab208026, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST. According to Uniprot annotation for HNRNPA1, isoform A1-A (34 kDa) is twenty times more abundant than isoform A1-B (39 kDa). The MW of HNRNPA1L2 is also 34 kD. In WB this antibody does not cross react with CCDC51 (expected MW 45 kDa).

All lanes:

Western blot - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] (<a href='/en-us/products/primary-antibodies/hnrnp-a1-citrulline-r92-antibody-epr20174-ab208026'>ab208026</a>) at 1/2000 dilution

Lanes 1 and 3:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a control vector containing GFP tag, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate at 20 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged PADI2 (WT) expression vector, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, whole cell lysate at 20 µg

Lane 4:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged PADI4 (WT) expression vector, treated with 10 mM calcium chloride and 10 µM Ionomycin for 2 hours, 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: 38 kDa

true

Exposure time: 5s

• Dot

Supplier Data

Dot Blot - Anti-hnRNP A1 (citrulline R92) antibody [EPR20174] - BSA and Azide free (AB251485)

This data was developed using ab208026, the same antibody clone in a different buffer formulation.

Dot blot analysis of hnRNP A1 (citrulline R92) labeled with ab208026 at 1/1000 dilution.

Lane 1 : hnRNP A1 (citrulline R92) peptide.

Lane 2 : hnRNP A1 non-citrulline peptide.

Lane 3 : hnRNP A3 (citrulline R113) peptide.

Lane 4 : hnRNP A0 (citrulline R85) peptide.

Lane 5 : CCDC51(citrulline R142) peptide.

Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

Based on sequence homology this antibody cross reacts with CCDC51 (citrulline R142) peptide (detected by dot blot only) and hnRNP A1L2 (citrulline R92) protein.

Exposure time : 3 minutes.

Blocking and dilution buffer : 5% NFDM/TBST.