Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Keap1 antibody [EPR22664-26] (ab227828)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22664-26] to Keap1
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-Keap1 antibody [EPR22664-26]
See all Keap1 primary antibodies
Description

Rabbit monoclonal [EPR22664-26] to Keap1
Host species

Rabbit
Tested Applications & Species

Application Species
IHC-P
Mouse

Human

WB
Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human lung tissue lysate; Wild-type HAP1, NK-92 and HepG2 whole cell lysates; mouse E14.5 liver tissue lysate. IHC-P: Human lung carcinoma tissue; mouse kidney tissue.
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.

Application	Species
IHC-P	Mouse Human
WB	Human

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: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22664-26
Isotype

IgG
Research areas

Western blot - Anti-Keap1 antibody [EPR22664-26] (ab227828)

All lanes : Anti-Keap1 antibody [EPR22664-26] (ab227828) at 1/1000 dilution

Lane 1 : Human lung tissue lysate
Lane 2 : Wild-type HAP1 whole cell lysate
Lane 3 : Keap1 knockout HAP1 whole cell lysate
Lane 4 : NK-92 (human malignant non-Hodgkin's lymphoma natural killer cell), whole cell lysate
Lane 5 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 6 : Mouse E14.5 liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 2-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 70 kDa
Observed band size: 70 kDa

Exposure time: 3 minutes

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

ab227828 was shown to specifically react with Keap1 in wild-type HAP1 cells as signal was lost in Keap1 knockout cells. Wild-type and Keap1 knockout samples were subjected to SDS-PAGE. ab227828 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 23242280, PMID: 20404090).

This blot was developed using a higher sensitivity ECL substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keap1 antibody [EPR22664-26] (ab227828)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Keap1 with ab227828 at 1/500 dilution (0.97 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on kidney tubules of mouse kidney (PMID: 28324005) is observed. The section was incubated with ab227828 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Keap1 antibody [EPR22664-26] (ab227828)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Keap1 with ab227828 at 1/500 dilution (0.97 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on tumor cells of human lung carcinoma (PMID: 20534738) is observed. The section was incubated with ab227828 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Anti-Keap1 antibody [EPR22664-26] (ab227828)

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