Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 09, 2021

Anti-REG4 antibody [EPR22810-327] (ab255820)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22810-327] to REG4
Suitable for: WB, IHC-P, IP
Reacts with: Human

Product name

Anti-REG4 antibody [EPR22810-327]
See all REG4 primary antibodies
Description

Rabbit monoclonal [EPR22810-327] to REG4
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	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: LoVo whole cell lysates; IHC-P: Human colon and colon carcinoma tissue; IP: LoVo 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

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR22810-327
Isotype

IgG
Research areas

Western blot - Anti-REG4 antibody [EPR22810-327] (ab255820)

All lanes : Anti-REG4 antibody [EPR22810-327] (ab255820) at 1/1000 dilution

All lanes : LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 18 kDa
Observed band size: 14 kDa
why is the actual band size different from the predicted?

Exposure time: 59 seconds

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20126989).

Negative control: HT-29 (PMID: 12455032).

Blocking/diluting buffer and concentration: 5% NFDM/TBST
Immunoprecipitation - Anti-REG4 antibody [EPR22810-327] (ab255820)

REG4 was immunoprecipitated from 0.35 mg LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10μg with ab255820 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab255820 1/1000 dilution (0.52 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate 10μg Lane 2: ab255820 IP in LoVo whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255820 in LoVo whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 min

This blot was developed using a higher sensitivity ECL substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-REG4 antibody [EPR22810-327] (ab255820)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling REG4 with ab255820 at 1/2000 dilution (0.262 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon carcinoma (PMID: 25295732) is observed. The section was incubated with ab255820 for 15 mins at RT. 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 20 mins.

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).
Western blot - Anti-REG4 antibody [EPR22810-327] (ab255820)

Anti-REG4 antibody [EPR22810-327] (ab255820) at 1/1000 dilution + Recombinant Human REG4 Protein at 20 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 18 kDa
Observed band size: 14 kDa why is the actual band size different from the predicted?

Exposure time: 59 seconds

Blocking/diluting buffer and concentration: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-REG4 antibody [EPR22810-327] (ab255820)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling REG4 with ab255820 at 1/2000 dilution (0.262 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon (PMID: 25295732) is observed. The section was incubated with ab255820 for 15 mins at RT. 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 20 mins.

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-REG4 antibody [EPR22810-327] (ab255820)

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