Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸ 10 µl 117 264 ₸

Order now and get it on Wednesday March 10, 2021

Anti-SEPP1 antibody [EPR24174-24] (ab277526)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR24174-24] to SEPP1
Suitable for: ICC, Flow Cyt, WB, IP
Reacts with: Human

Product name

Anti-SEPP1 antibody [EPR24174-24]
See all SEPP1 primary antibodies
Description

Rabbit monoclonal [EPR24174-24] to SEPP1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC	Human
IP	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 liver tissue lysate; Human plasma; HepG2 whole cell lysate. ICC: HepG2 cells. Flow cyt: HepG2 cells. 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

EPR24174-24
Isotype

IgG

Immunocytochemistry - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 cells labelling SEPP1 with ab277526 at 1/50 (12.3 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.
Flow Cytometry - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling SEPP1 with ab277526 at 1/50 dilution (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.
Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

All lanes : Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution

Lane 1 : Human liver tissue lysate
Lane 2 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

Predicted band size: 43 kDa
Observed band size: 50 kDa
why is the actual band size different from the predicted?

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

The observed MW is consistent with what has been described in the literature (PMID:19345254, 27859223).

Low expression: Human skeletal muscle(PMID:23038251).

Exposure time: 3 minutes.
Immunoprecipitation - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

SEPP1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate with ab277526 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277526 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: ab277526 IP in HepG2 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab277526 in HepG2 whole cell lysate

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

Exposure time: 3 minutes.
Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution + Human plasma at 10 µl

Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 43 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

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

The observed MW is consistent with what has been described in the literature (PMID:19345254, 27859223).

Exposure time: 10 seconds.
Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

All lanes : Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution

Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell) (treated with 300ng/ml Brefeldin A (BFA) for 24 hours) whole cell lysate
Lane 2 : HepG2 (treated with 300ng/ml Brefeldin A (BFA) for 24 hours) whole cell lysate treated with PNGase F

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 43 kDa
Observed band size: 35-47,50-60 kDa why is the actual band size different from the predicted?

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

The observed MW is consistent with what has been described in the literature (PMID:19345254, 27859223).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time: 3 minutes.
Anti-SEPP1 antibody [EPR24174-24] (ab277526)

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