Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-Angiotensinogen antibody [EPR24118-2]
See all Angiotensinogen primary antibodies
Description

Rabbit monoclonal [EPR24118-2] to Angiotensinogen
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IHC-P	Rat Human
IP	Human
WB	Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human plasma, lung, liver and kidney tissue lysates; Rat plasma; Rat lung, kidney and liver tissue tissue lysate; HepG2 and Huh7 whole cell lysates. Flow cyt: HepG2 cells. ICC: HepG2 cells. IHC-P: Rat kidney tissue; Human placenta, liver, and kidney tissue. IP: HepG2 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24118-2
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat kidney (PMID: 28716988). The section was incubated with ab276132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Immunocytochemistry - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized HepG2 cells labelling Angiotensinogen with ab276132 at 1/100 (4.27 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.
Western blot - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate
Lane 2 : Huh7 (human hepatocellar carcinoma epithelial cell), whole cell lysate
Lane 3 : Human lung tissue lysate
Lane 4 : Human liver tissue lysate
Lane 5 : Human kidney tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 53 kDa
Observed band size: 48-70 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what has been described in literature(PMID: 9694881).

Lanes 1 & 2 of the blot were developed using a higher sensitivity ECL substrate.

Exposure times: Lane 1, 2: 59 seconds; Lane 3: 3.25 seconds;Lane 4, 5: 15 seconds
Flow Cytometry - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Angiotensinogen with ab276132 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Angiotensinogen was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate with ab276132 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab276132 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

Lane 2: ab276132 IP in HepG2 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab276132 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 23 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human placenta (PMID: 21215447). The section was incubated with ab276132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Western blot - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

Lane 1 : Rat lung tissue lysate
Lane 2 : Rat liver tissue lysate
Lane 3 : Rat kidney tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 53 kDa
Observed band size: 48-70 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what have been described in literature(PMID: 9694881).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time: 127 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human kidney. The section was incubated with ab276132 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Western blot - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

Lane 1 : Human plasma
Lane 2 : Rat plasma

Lysates/proteins at 20 µl per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Predicted band size: 53 kDa
Observed band size: 48-70 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what have been described in literature(PMID: 9694881).

Exposure times:

Lane 1: 15 seconds
Lane 2: 3.25 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human liver. The section was incubated with ab276132 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Anti-Angiotensinogen antibody [EPR24118-2] (ab276132)