Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / NAMALWA (human Burkitt's lymphoma B lymphocyte, Right) cells labelling AICDA with ab269454 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Blac) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : K-562 cell line (PMID : 27217538).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling AICDA with ab269454 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in germinal center cells of human tonsil (PMID : 23877718, 15732141, PMID : 29251015).

The section was incubated with ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Immunohistochemical analysis of paraffin-embedded Human Hodgkin lymphoma tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in part of tumor cells of human Hodgkin lymphoma (PMID : 15732141).

The section was incubated with ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma tissue labeling AICDA with ab269454 at 1/4000 dilution (0.12ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Mainly cytoplasmic staining (weak nuclear staining) in part of tumor cells of human diffuse large B-cell lymphoma (PMID : 29251015).

The section was incubated with ab255611 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• IP

Unknown

Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

AICDA was immunoprecipitated from 0.35 mg NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab269454 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269454 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : NAMALWA whole cell lysate 10ug.

Lane 2 : ab269454 IP in NAMALWA whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269454 in NAMALWA whole cell lysate.

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

Exposure time : 90 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

All lanes:

Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - ChIP Grade (<a href='/en-us/products/primary-antibodies/aicda-antibody-epr23436-45-chip-grade-ab269454'>ab269454</a>)

Predicted band size: 23 kDa

Observed band size: 24 kDa

false

• ChIP

Unknown

ChIP - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

Chromatin was prepared from Ramos cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

The ChIP was performed with 25 μg of chromatin, 5 μg of ab269454 (red), or 5 μg of rabbit normal IgG ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

Primers and probes are commercial primers from paper : PMC2905439

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

• IP

Unknown

Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

AICDA was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab269454 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab269454 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Ramos whole cell lysate 10ug.

Lane 2 : ab269454 IP in Ramos whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab269454 in Ramos whole cell lysate.

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

Exposure time : 90 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454).

All lanes:

Immunoprecipitation - Anti-AICDA antibody [EPR23436-45] - ChIP Grade (<a href='/en-us/products/primary-antibodies/aicda-antibody-epr23436-45-chip-grade-ab269454'>ab269454</a>)

Predicted band size: 23 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-AICDA antibody [EPR23436-45] - ChIP Grade - BSA and Azide free (AB269457)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab269454). False colour image of Western blot : Anti-AICDA antibody [EPR23436-45] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab269454 was shown to bind specifically to AICDA. A band was observed at 22 kDa in wild-type Raji cell lysates with no signal observed at this size in AICDA knockout cell line ab277185 (knockout cell lysate ab277227). To generate this image, wild-type and AICDA knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-AICDA antibody [EPR23436-45] - ChIP Grade (<a href='/en-us/products/primary-antibodies/aicda-antibody-epr23436-45-chip-grade-ab269454'>ab269454</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

AICDA knockout Raji cell lysate at 20 µg

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 22 kDa

false