Anti-Matrin 3 antibody [EPR10635(B)]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

ab151714 was shown to react with MATR3 in wild-type HAP1 cells in immunocytochemistry with loss of signal observed in a MATR3 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab151714 at dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 µg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Intracellular Flow Cytometry analysis of 4%paraformaldehyde fixedHepG2 (human hepatocellular carcinoma) cells labeling Matrin 3 with purified ab151714 at a dilution of 1/220. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin embedded Human Ovarian carcinoma tissue using unpurified ab151714 showing +ve staining.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin-embedded human colon tissue sections labeling Matrin 3 with purified ab151714 at a dilution of 1/2000 (0.7 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using citrate Buffer, pH 6.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin embedded Human Lung carcinoma tissue using unpurified ab151714 showing +ve staining.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling Matrin 3 with unpurified ab151714 at 1/250 dilution.

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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Matrin 3 with purified ab151714 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor^® 594) at 1/200. DAPI (blue) was used as a nuclear counterstain. Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin embedded normal Human uterus tissue using unpurified ab151714 showing +ve staining.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Matrin 3 with unpurified ab151714 at 1/250 dilution.

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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunofluorescent analysis of HepG2 cells labeling Matrin 3 with unpurified ab151714 at 1/250 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of Paraffin-embedded rat liver tissue sections labeling Matrin 3 with purified ab151714 at a dilution of 1/2000 (0.7 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using citrate Buffer, pH 6.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue sections labeling Matrin 3 with purified ab151714 at a dilution of 1/2000 (0.7 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using citrate Buffer, pH 6.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• WB

Collaborator

Western blot - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

ab151714 was shown to react with MATR3 in wild-type HAP1 cells in Western blot with loss of signal observed in a MATR3 knockout cell line. Wild-type HAP1 and MATR3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab151714 overnight at 4 °C at a 1/10000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2 μg/ml before imaging. These data were 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-Matrin 3 antibody [EPR10635(B)] (ab151714)

Predicted band size: 95 kDa

false

• WB

Unknown

Western blot - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

Blocking/Diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-Matrin 3 antibody [EPR10635(B)] (ab151714) at 1/10000 dilution

Lane 1:

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

Lane 2:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Rat brain tissue 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 1/20000 dilution

Predicted band size: 59 kDa,95 kDa

Observed band size: 125 kDa

false

• WB

Unknown

Western blot - Anti-Matrin 3 antibody [EPR10635(B)] (AB151714)

All lanes:

Western blot - Anti-Matrin 3 antibody [EPR10635(B)] (ab151714) at 1/1000 dilution

Lane 1:

HEK293T cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 95 kDa

false