Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

Immunohistochemical analysis of paraffin-embedded A : Wild-type A549 (Human lung carcinoma epithelial cell) cell pellet, B : TRIM21 knockout A549 (ab267080) cell pellet labelling TRIM21/SS-A with ab207728 at 1/500 dilution (1.212 μg/ml) followed by LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody at a ready to use concentration. Positive staining on (A) wild-type A549 cell pellet, no staining on (B) TRIM21 knockout A549 (ab267080) cell pellet. The section was incubated with ab207728 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

This data was developed using ab207728, the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

Immunohistochemistry analysis of paraffin-embedded human tonsil tissue sections labelling TRIM21/SS-A with ab207728 at 1/100 dilution. The section was incubated with ab207728 for 10 mins at room temperature. Ready to use Leica DS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Positive staining on human tonsil. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using ab207728, the same antibody clone in a different buffer formulation.

• WB

Supplier Data

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

Blocking/Dilution buffer : 5% NFDM/TBST.

The level of TRIM21 expression can be elevated by IFN alpha treatment (PMID : 18071879).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207728).

All lanes:

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (ab232549) at 1/1000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 10 µg

Lane 2:

HeLa whole cell lysate treated with 10 ng/ml human interferon-α (<a href='/en-us/products/proteins-peptides/recombinant-human-interferon-alpha-1-protein-active-ab48750'>ab48750</a>) for 16 hours 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 1/10000 dilution

Predicted band size: 54 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

This data was developed using ab207728, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab207728 Anti-TRIM21/SS-A antibody [EPR20290] was shown to specifically react with TRIM21/SS-A in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267025 (knockout cell lysate ab257767) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. ab207728 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/500 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

TRIM21 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM21 (SS-A) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trim21-ss-a-knockout-a549-cell-line-ab267025'>ab267025</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 54 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

This data was developed using ab207728, the same antibody clone in a different buffer formulation.

ab207728 was shown to react with TRIM21 in wild-type A549 cells in Western blot with loss of signal observed in TRIM21 knockout cell line ab267025. Wild-type A549 and TRIM21 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab207728 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 lysate at 30 µg

Lane 2:

TRIM21 knock-out A549 lysate at 30 µg

false

• WB

Lab

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

This data was developed using ab207728, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab207728 Anti-TRIM21/SS-A antibody [EPR20290] was shown to specifically react with TRIM21/SS-A in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267024 (knockout cell lysate ab257766) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. ab207728 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TRIM21 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TRIM21 (SS-A) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trim21-ss-a-knockout-a549-cell-line-ab267024'>ab267024</a>)

Lane 3:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 4:

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

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 54 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (AB232549)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207728).

Lanes 1 - 4 : Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab207728 was shown to specifically react with in wild-type HAP1 cells as signal was lost in TRIM21 knockout cells. Wild-type and TRIM21 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. ab207728 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-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

TRIM21 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

MOLT-4 whole cell lysate at 20 µg

Predicted band size: 54 kDa

false