重组Anti-Galectin 1抗体[EPR3206(2)]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling Galectin 1 with Purified ab138513 at 1 : 30 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

ab138513 was shown to react with LGALS1 in wild-type HAP1 cells in immunocytochemistry with loss of signal observed in a LGALS1 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 ab138513 at 1/250 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Galectin 1 with Purified ab138513 at 1/16000 dilution (0.02 µg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. M3

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

Immunocytochemistry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling Galectin 1 with Purified ab138513 at 1/50 dilution (5.4 µg/mL). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

ab138513 was shown to react with LGALS1 in wild-type HAP1 cells in Western blot with loss of signal observed in a LGALS1 knockout cell line. Wild-type HAP1 and LGALS1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab138513 overnight at 4 °C at a 1/10000 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-Galectin 1 antibody [EPR3206(2)] (ab138513) at 1/10000 dilution

Lane 1:

Wild-type HAP1 lysate at 10 µg

Lane 2:

LGALS1 knock-out HAP1 lysate at 10 µg

Observed band size: 14 kDa

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• WB

Lab

Western blot - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

Lanes 1- 2 : Merged signal (red and green). Green - ab138513 observed at 15 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab138513 was shown to react with Galectin 1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266850 (knockout cell lysate ab256974) was used. Wild-type HEK-293T and LGALS1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab138513 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 Dilution and a 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-Galectin 1 antibody [EPR3206(2)] (ab138513) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

LGALS1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LGALS1 (Galectin 1) knockout HEK-293T cell line (<a href='/products/cell-lines/human-lgals1-galectin-1-knockout-hek-293t-cell-line-ab266850'>ab266850</a>)

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

Lanes 1 - 2 : Merged signal (red and green). Green - ab138513 observed at 15 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab138513 was shown to specifically react with Galectin 1 in wild-type HAP1 cells as signal was lost in LGALS1 (Galectin 1) knockout cells. Wild-type and LGALS1 (Galectin 1) knockout samples were subjected to SDS-PAGE. ab138513 and ab9484 (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-Galectin 1 antibody [EPR3206(2)] (ab138513) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

LGALS1 (Galectin 1) knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 15 kDa

false

• WB

Unknown

Western blot - Anti-Galectin 1 antibody [EPR3206(2)] (AB138513)

All lanes:

Western blot - Anti-Galectin 1 antibody [EPR3206(2)] (ab138513) at 1/10000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

Lane 2:

Mouse heart lysate at 15 µg

Lane 3:

Rat heart lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 14 kDa

false