Antibodies, Proteins, Kits and Reagents for Life Science

345 091 ₸ Product size

100 µl 345 091 ₸ 10 µl 140 716 ₸

Order now and get it on Tuesday March 02, 2021

Anti-IL-12A antibody [EP5737] (ab131039)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP5737] to IL-12A
Suitable for: WB, IHC-P, Flow Cyt
Reacts with: Mouse, Human

Product name

Anti-IL-12A antibody [EP5737]
See all IL-12A primary antibodies
Description

Rabbit monoclonal [EP5737] to IL-12A
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human IL-12A aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
Positive control
- NIH3T3, HuT-78, and Raw 264.7 cell lysates; Human kidney tissue, Human gastric carcinoma, Human colon tissue.
General notes

Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP5737
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)

Immunohistochemical staining of paraffin embedded human gastric carcinoma tissue section labelling IL-12A with purified ab131039 at dilution of 1/800. The secondary antibody used was HRP conjugated Goat Anti-Rabbit IgG H&L (ab97051) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Flow Cytometry - Anti-IL-12A antibody [EP5737] (ab131039)

Flow cytometry analysis of Raw264.7 cells treated with 100ng/ml for LPS 24hour cells labelling IL-12A with ab131039 at 1/19000 dilution (1.02 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).
Flow Cytometry - Anti-IL-12A antibody [EP5737] (ab131039)

Flow cytometry analysis of THP-1 cells treated with 100ng/ml PMA for 30min,then 1µg/ml LPS for 24hour cells labelling IL-12A with ab131039 at 1/19000 dilution (1.02 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Dylight488® (Ab98462) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue). Untreated cells (Green).
Western blot - Anti-IL-12A antibody [EP5737] (ab131039)

Anti-IL-12A antibody [EP5737] (ab131039) at 1/1000 dilution (purified) + HuT-78 (human Sezary syndrome) whole cell lysate at 20 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer 5% NFDM/TBST
Western blot - Anti-IL-12A antibody [EP5737] (ab131039)

All lanes : Anti-IL-12A antibody [EP5737] (ab131039) at 1/5000 dilution (purified)

Lane 1 : NIH/3T3 (mouse embryo) whole cell lysate
Lane 2 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 25 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer 5% NFDM/TBST
Western blot - Anti-IL-12A antibody [EP5737] (ab131039)

All lanes : Anti-IL-12A antibody [EP5737] (ab131039) at 1/1000 dilution (unpurified)

Lane 1 : NIH3T3 cell lysate
Lane 2 : HuT-78 cell lysate
Lane 3 : Raw 264.7 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 25 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)

Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labelling IL-12A with unpurified ab131039 at dilution of 1:250.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039) This image is courtesy of an anonymous Abreview.

Formaldehyde-fixed, paraffin-embedded human colon tissue stained for IL-12A using ab131039 at 1/500 dilution in immunohistochemical analysis.
See Abreview
Anti-IL-12A antibody [EP5737] (ab131039)

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