We have recently purchased mouse monoclonal anti-PDI antibody RL90 (cat# NB300-517). From the information available, it appears that this was generated with purified rat PDI serving as immunogen. We would like to know whether there is any information available on the epitope in the PDI molecule recognized by the RL90 antibody? I could not find this information in the description of the antibody.

Name: Protein Disulfide Isomerase/P4HB Antibody (RL90)
Brand: Novus Biologicals
SKU: NB300-517
Rating: 5 (3 reviews)

Yes, you are correct: purified rat PDI protein was used as the immunogen for NB300-517, PDI Antibody (RL90). We do not normally epitope map our antibodies, so I think it is unlikely that we know where on the PDI protein this antibody binds. The lab have now confirmed to me that no epitope mapping been carried out for NB300-517, and hence we do not have any additional detail on the binding region of this antibody.

Protein Disulfide Isomerase/P4HB Antibody (RL90)

Novus Biologicals, part of Bio-Techne | Catalog # NB300-517

(3)

Citations (29)

Product Datasheet / COA / SDS

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Product Summary
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Independent Antibodies

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Hamster, Monkey, Primate, Rabbit

Cited:

Human, Mouse, Rat, Primate, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Block/Neutralize, ELISA, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunomicroscopy, Immunoprecipitation, Knockdown Validated, Western Blot

Cited:

Block/Neutralize, Flow Cytometry, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2A Clone # RL90

Concentration

1 mg/ml

View all Protein Disulfide Isomerase/P4HB Antibodies »

Product Summary for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Immunogen

Purified rat Protein Disulfide Isomerase/P4HB protein.

Reactivity Notes

Rabbit reactivity reported in scientific literature (PMID: 16150729). Bovine (PMID: 14592831). Primate (PMID: 20667829). Monkey reactivity reported in scientific literature (PMID: 27487388). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Specificity

NB300-517 detects Protein Disulfide Isomerase/P4HB from human, rat, porcine and mouse tissues as well as hamster cells.

Marker

Endoplasmic Reticulum Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of Protein Disulfide Isomerase/P4HB was performed by loading 25 ug of HepG2 (Lane 1), Hela (Lane 2) and NIH-3T3 (Lane 3) cell lysates.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining of Phalloidin (purple) and PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 650 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Both normal and cancer biopsies of deparaffinized Human colon tissue tissues.

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of Hela cells compared to an isotype control (blue).

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of PDIA2 expression in 1) human brain and 2) rat liver tissues extracts, 3) HeLa and 4) Hek 293 whole cell lysates.

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cell surface Protein Disulfide Isomerase/P4HB associated with GC-AC. Proteins were pulled down with anti GC-A antibody from HUVEC or HMC membrane preparations and detected with anti-Protein Disulfide Isomerase/P4HB antibody (RL90)compared to control IP (no anti GC-A) and non-immunoprecipitated proteins from HMC and HUVEC. Image collected and cropped by CiteAb from the following publication (http://dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in p19 Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with Dylight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with Dylight 350 Phalloidin at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (red) were stained with DRAQ5 at a concentration of 1ug/ml for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 550 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 488 Phalloidin at a dilution of 1:300 (1 unit/ml final concentration) for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in A549 Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in HMVEC Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in NS-1 Cells.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cancer biopsies of deparaffinized Human lung adenocarcinoma tissues.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Both normal and cancer biopsies of deparaffinized Human tonsil tissues.

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of NIH/3T3 cells compared to an isotype control (blue).

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of K562 cells compared to an isotype control (blue).

Knockdown Validated: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Down regulated PDIA2 attenuates CNP-mediated cGMP activation in HMCs. Western blotting with RL90 antibody shows partial knockdown of Protein Disulfide Isomerase/P4HB in HMCs.

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Applications for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Application

Recommended Usage

ELISA

1:100 - 1:2000

Flow Cytometry

1:10 - 1:1000

Immunocytochemistry/Immunofluorescence

1:100

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Frozen

1:100

Immunohistochemistry-Paraffin

1:100

Immunomicroscopy

1:10 - 1:500

Immunoprecipitation

1:10 - 1:500

Western Blot

1:1000

Application Notes

IHC usage was reported in scientific literature (see Tanaka S, et al). ELISA assay reported in literature (see Kaetzel CS, et al). Flow Cytometry usage was reported in scientific literature (see Ryser HJ, et al). Blocking/Neutralization usage was reported in the literature (PMID:12791669 and PMID: 22013897)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 3 reviews rated 5 using NB300-517 in the following applications:

Western Blot (3 Reviews)

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS with 1 mg/ml BSA

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: Protein Disulfide Isomerase/P4HB

P4HB, the beta subunit of prolyl 4-hydroxylase, a highly abundant member of the PDI family. P4HB is a multifunctional enzyme, involved in hydroxylation of prolyl residues in preprocollagen, as well as continual rearrangement of disulfide bonds. Other known functions include its ability to act as a chaperone that inhibits aggregation of misfolded proteins in a concentration-dependent manner, its ability to bind thyroid hormone, its role in both the influx and efflux of S-nitrosothiol-bound nitric oxide, and its function as a subunit of the microsomal triglyceride transfer protein complex.

Long Name

Proline 4-Hydroxylase, Beta Polypeptide

Alternate Names

DSI, ERBA2L, GIT, PDI, PDIA1, PO4DB, PO4HB, PROHB, Thbp

Gene Symbol

P4HB

Additional Protein Disulfide Isomerase/P4HB Products

Product Documents for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for Protein Disulfide Isomerase/P4HB Antibody (RL90)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations
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FAQs

Protein Disulfide Isomerase/P4HB Antibody (RL90)

Novus Biologicals, part of Bio-Techne | Catalog # NB300-517

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications

Validated:

Cited:

Label

Antibody Source

Concentration

View all Protein Disulfide Isomerase/P4HB Antibodies »

Product Summary for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Immunogen

Reactivity Notes

Specificity

Marker

Clonality

Host

Isotype

Scientific Data Images for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Applications for Protein Disulfide Isomerase/P4HB Antibody (RL90)

ELISA

Flow Cytometry

Immunocytochemistry/Immunofluorescence

Immunohistochemistry

Immunohistochemistry-Frozen

Immunohistochemistry-Paraffin

Immunomicroscopy

Immunoprecipitation

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Purification

Formulation

Preservative

Concentration

Shipping

Stability & Storage

Background: Protein Disulfide Isomerase/P4HB

Long Name

Alternate Names

Gene Symbol

Additional Protein Disulfide Isomerase/P4HB Products

Product Documents for Protein Disulfide Isomerase/P4HB Antibody (RL90)

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for Protein Disulfide Isomerase/P4HB Antibody (RL90)