Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR5299] to HPRT
Suitable for: WB, IP, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-HPRT antibody [EPR5299]
See all HPRT primary antibodies
Description

Rabbit monoclonal [EPR5299] to HPRT
Host species

Rabbit
Specificity

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Tested Applications & Species

Application	Species
IHC-P	Human
IP	Rat
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa HepG2, A549, and MCF7 cell lysates, Mouse brain and Rat brain tissue lysates IHC-P:Human lung and colon cancer IP: Rat brain 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.
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 long term. Avoid freeze / thaw cycle.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR5299
Isotype

IgG
Research areas

Western blot - Anti-HPRT antibody [EPR5299] (ab109021)

All lanes : Anti-HPRT antibody [EPR5299] (ab109021) at 0.02 µg/ml (purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Mouse brain lysates
Lane 3 : Rat brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.02 µg/ml

Predicted band size: 25 kDa

Blocking and diluting buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling HPRT with Purified ab109021 at 1:150 dilution (1.19 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control:PBS instead of the primary antibody.Hematoxylinwas used as a counterstain
Immunoprecipitation - Anti-HPRT antibody [EPR5299] (ab109021)

ab109021 (purified) at 1:30 dilution (2µg) immunoprecipitating HPRT in Rat brain lysate.
Lane 1 (input): Rat brain lysate 10µg
Lane 2 (+): ab109021 & Rat brain lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109021 in Rat brain lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Western blot - Anti-HPRT antibody [EPR5299] (ab109021)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HPRT1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A431 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab109021 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

Unpurified ab109021 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were examined. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab109021 and ab8245 (Mouse anti GAPDH loading control) were both diluted at 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-HPRT antibody [EPR5299] (ab109021)

All lanes : Anti-HPRT antibody [EPR5299] (ab109021) at 1/10000 dilution (Unpurified)

Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : MCF7 cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 25 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (ab109021)

Unpurified ab109021 at 1/50 dilution staining HPRT in Human colon by Immunohistochemistry, Paraffin-embedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-HPRT antibody [EPR5299] (ab109021)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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