Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

Order now and get it on Tuesday March 09, 2021

Anti-CYP27B1 antibody [EPR20271] (ab206655)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20271] to CYP27B1
Suitable for: WB, IHC-P, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-CYP27B1 antibody [EPR20271]
See all CYP27B1 primary antibodies
Description

Rabbit monoclonal [EPR20271] to CYP27B1
Host species

Rabbit

Tested Applications & Species

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

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HaCaT, MeWo and HeLa whole cell lysates; Human fetal heart, fetal brain, fetal kidney and fetal liver lysates; Mouse heart and kidney lysates; Rat heart, kidney and brain lysates. IHC-P: Human, mouse and rat kidney tissues. IP: MeWo 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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20271
Isotype

IgG

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Strong cytoplasmic staining on distal convoluted tubules of human kidney. Weaker cytoplasmic staining on proximal convoluted tubules [PMID: 23416105] [PMID: 12050281].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse kidney tubules [PMID: 23416105] [PMID: 12050281].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)

All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/2000 dilution

Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 2 : HaCaT (Human keratinocyte cell line) whole cell lysate
Lane 3 : MeWo (Human malignant melanoma cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : Human fetal heart lysate
Lane 6 : Human fetal brain lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 5-6 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 57 kDa
Observed band size: 45,70 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

THP-1 is a negative control (PMID: 16930540).

The molecular weight observed is consistent with what has been described in the literature (PMID: 16061850).
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)

All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/1000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 57 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16061850).
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)

All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/1000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat heart lysate
Lane 4 : Rat kidney lysate
Lane 5 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 57 kDa
Observed band size: 57 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1, 2, 3 and 4: 1 second; Lane 5: 3 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat kidney tubules [PMID: 23416105] [PMID: 12050281].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-CYP27B1 antibody [EPR20271] (ab206655)

CYP27B1 was immunoprecipitated from 0.35 mg of MeWo (Human malignant melanoma cell line) whole cell lysate with ab206655 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab206655 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: MeWo whole cell lysate, 10 μg (Input).

Lane 2: ab206655 IP in MeWo whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206655 in MeWo whole cell lysate.

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

Exposure time: 30 seconds.
Anti-CYP27B1 antibody [EPR20271] (ab206655)

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