Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 09, 2021

Anti-AACT antibody [EPR17088-68] (ab205198)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17088-68] to AACT
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Human

Product name

Anti-AACT antibody [EPR17088-68]
See all AACT primary antibodies
Description

Rabbit monoclonal [EPR17088-68] to AACT
Host species

Rabbit
Specificity

This antibody recognizes a different epitope of AACT compared to ab204514.

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human plasma (untreated and PNGase F treated) and blood; Human fetal spleen lysate. IHC-P: Human tonsil tissue. ICC/IF: Raji cells. IP: Human plasma 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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17088-68
Isotype

IgG
Research areas

Western blot - Anti-AACT antibody [EPR17088-68] (ab205198)

Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution + Human plasma at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 47 kDa
Observed band size: 50-70 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 9880565).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AACT antibody [EPR17088-68] (ab205198)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling AACT with ab205198 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab205198) at 1/500 dilution. Cytoplasmic staining on human tonsil tissue 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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-AACT antibody [EPR17088-68] (ab205198)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling AACT with ab205198 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining on Raji cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab205198 at 1/50 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Western blot - Anti-AACT antibody [EPR17088-68] (ab205198)

Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + His-Tagged Human SERPINA3 (aa 24 to 324) recombinant protein 100ng

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 47 kDa
Observed band size: 33 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Diluting buffer and concentration 5% NFDM/TBST
Western blot - Anti-AACT antibody [EPR17088-68] (ab205198)

Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human blood at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-AACT antibody [EPR17088-68] (ab205198)

Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human fetal spleen lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-AACT antibody [EPR17088-68] (ab205198)

All lanes : Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution

Lane 1 : Untreated Human plasma
Lane 2 : Human plasma treated with PNGase F

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 47 kDa
Observed band size: 50-70 kDa why is the actual band size different from the predicted?

Exposure time: 4 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

De-glycosylation treatment with PNGase, demonstrates a molecular weight shift from 50-70kDa to 46kDa. This result is consistent with what has been described in literature the literature (PMID:) 9880565.
Immunoprecipitation - Anti-AACT antibody [EPR17088-68] (ab205198)

AACT was immunoprecipitated from 1 mg of Human plasma lysate with ab205198 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab205198 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

Lane 1: Human plasma lysate, 10 μg (Input).

Lane 2: ab205198 IP in Human plasma lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205198 in Human plasma lysate.

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

Exposure time: 30 seconds.
Anti-AACT antibody [EPR17088-68] (ab205198)

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