Anti-Granulin 抗体 [EPR18539-59] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Granulin antibody [EPR18539-59] - BSA and Azide free (AB227816)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage cell line) cells labeling Granulin with ab187070 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on J774A.1 cell line.

The nuclear counter statin is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Negative control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187070).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Granulin antibody [EPR18539-59] - BSA and Azide free (AB227816)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cell line labeling Granulin with ab187070 at 1/1000 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

Goat Anti-Rabbit IgG H&L (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 (ab187070).

• IP

Supplier Data

Immunoprecipitation - Anti-Granulin antibody [EPR18539-59] - BSA and Azide free (AB227816)

Granulin was immunoprecipitated from 0.35 mg of J774A.1 (mouse reticulum cell sarcoma monocyte macrophage cell line) whole cell lysate with ab187070 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab187070 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : J774A.1 lysate 10 μg (Input).

Lane 2 : ab187070 IP in J774A.1 lysate (+).

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab187070 in J744A.1 whole cell lysate (-).

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187070).

All lanes:

Immunoprecipitation - Anti-Granulin antibody [EPR18539-59] (<a href='/products/primary-antibodies/granulin-antibody-epr18539-59-ab187070'>ab187070</a>)

Predicted band size: 64 kDa

Observed band size: 88 kDa

true

• WB

Lab

Western blot - Anti-Granulin antibody [EPR18539-59] - BSA and Azide free (AB227816)

This data was developed using the same antibody clone in a different buffer formulation (ab208777)

ab208777 was shown to react with GRN in wild-type HEK293T cells in Western blot with loss of signal observed in GRN knockout cell line ab266738. Wild-type HEK293T and GRN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab208777 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2ug/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-Granulin antibody [EPR15864] (<a href='/products/primary-antibodies/granulin-antibody-epr15864-ab208777'>ab208777</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T lysate at 20 µg

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell line (<a href='/products/cell-lines/human-grn-granulin-knockout-hek-293t-cell-line-ab266738'>ab266738</a>)

Lane 2:

Western blot - Human GRN (Granulin) knockout HEK-293T cell lysate (<a href='/products/cell-lysates/human-grn-granulin-knockout-hek-293t-cell-lysate-ab257235'>ab257235</a>)

Observed band size: 64 kDa

false