Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling alpha 1 Antitrypsin with ab207303 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human liver is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207303).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling alpha 1 Antitrypsin with ab207303 at 1/50 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 K562 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab207303 at 1/50 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) 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 (ab207303).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling alpha 1 Antitrypsin with ab207303 at 1/50 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 HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse Alexa Fluor^® 594 (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab207303 at 1/50 dilution, followed by Goat Anti-Mouse Alexa Fluor^® 594 (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) seconday 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 (ab207303).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling alpha 1 Antitrypsin with ab207303 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on cancer cells of human hepatocellular carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207303).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

alpha 1 Antitrypsin was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab207303 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab207303 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : HepG2 whole cell lysate 10ug (Input).
Lane 2 : ab207303 IP in HepG2 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207303 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207303).

All lanes:

Immunoprecipitation - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] (<a href='/ko/products/primary-antibodies/alpha-1-antitrypsin-antibody-epr17087-50-ab207303'>ab207303</a>)

Predicted band size: 46 kDa

false

• WB

Collaborator

Western blot - Anti-alpha 1 Antitrypsin antibody [EPR17087-50] - BSA and Azide free (AB240375)

This data was developed using ab207303, the same antibody clone in a different buffer formulation.

ab207303 was shown to react with SERPINA1 in wild-type HepG2 cells in Western blot with loss of signal observed in a SERPINA1 knockout cell line. Wild-type HepG2 and SERPINA1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab207303 overnight at 4 °C at a 1/5000 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-alpha 1 Antitrypsin antibody [EPR17087-50] (<a href='/ko/products/primary-antibodies/alpha-1-antitrypsin-antibody-epr17087-50-ab207303'>ab207303</a>) at 1/5000 dilution

Lane 1:

Wild-type HepG2 lysate at 30 µg

Lane 2:

SERPINA1 knock-out HepG2 lysate at 30 µg

false