概述

  • 产品名称
    Anti-P4HB抗体[RL90]
    参阅全部 P4HB 一抗
  • 描述
    小鼠单克隆抗体[RL90] to P4HB
  • 宿主
    Mouse
  • 经测试应用
    适用于: ICC/IF, Electron Microscopy, IHC-P, IHC-Fr, IP, WB, ELISA, Inhibition Assay, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Dog, Human, Pig, Monkey, African green monkey
    预测可用于: Drosophila melanogaster
  • 免疫原

    Other Immunogen Type corresponding to Rat P4HB. Purified rat PDIA1/P4HB protein.
    Database link: P04785

  • 阳性对照
    • rat liver

性能

应用

Our Abpromise guarantee covers the use of ab2792 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/100. PubMed: 17308099
Electron Microscopy Use at an assay dependent concentration. PubMed: 21886772
IHC-P 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr 1/100.
IP Use at an assay dependent concentration. This antibody has been shown to inhibit the activity of PDI in vitro. It has also been found to inhibit disulfide bond reduction of the HIV protein, gp120, at the cell surface of CHO cells and human lymphoid cells.
WB 1/1000. Detects a band of approximately 59-61 kDa (predicted molecular weight: 58 kDa). If there is no signal or signal is weak, more concentrated antibody could be used in addition to using less stringent blocking conditions (e.g., BSA instead of milk, incubating the antibody in PBST or TBST only, lower milk percentage).
ELISA Use at an assay dependent concentration.
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

靶标

  • 功能
    This multifunctional protein catalyzes the formation, breakage and rearrangement of disulfide bonds. At the cell surface, seems to act as a reductase that cleaves disulfide bonds of proteins attached to the cell. May therefore cause structural modifications of exofacial proteins. Inside the cell, seems to form/rearrange disulfide bonds of nascent proteins. At high concentrations, functions as a chaperone that inhibits aggregation of misfolded proteins. At low concentrations, facilitates aggregation (anti-chaperone activity). May be involved with other chaperones in the structural modification of the TG precursor in hormone biogenesis. Also acts a structural subunit of various enzymes such as prolyl 4-hydroxylase and microsomal triacylglycerol transfer protein MTTP.
  • 序列相似性
    Belongs to the protein disulfide isomerase family.
    Contains 2 thioredoxin domains.
  • 细胞定位
    Endoplasmic reticulum lumen. Melanosome. Cell membrane. Highly abundant. In some cell types, seems to be also secreted or associated with the plasma membrane, where it undergoes constant shedding and replacement from intracellular sources (Probable). Localizes near CD4-enriched regions on lymphoid cell surfaces. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Cellular thyroid hormone binding protein antibody
    • Cellular thyroid hormone-binding protein antibody
    • Collagen prolyl 4 hydroxylase beta antibody
    • Disulphide Isomerase antibody
    • DSI antibody
    • EC 5.3.4.1 antibody
    • Endoplasmic reticulum resident protein 59 antibody
    • ER protein 59 antibody
    • ERBA2L antibody
    • ERp59 antibody
    • GIT antibody
    • Gltathione insulin transhydrogenase antibody
    • Glutathione insulin transhydrogenase antibody
    • P4HB antibody
    • P4Hbeta antibody
    • p55 antibody
    • PDI antibody
    • PDIA1 antibody
    • PDIA1_HUMAN antibody
    • PDIR antibody
    • PHDB antibody
    • PO4DB antibody
    • PO4HB antibody
    • Procollagen proline 2 oxoglutarate 4 dioxygenase (proline 4 hydroxylase) beta polypeptide (protein disulfide isomerase associated 1) antibody
    • Procollagen proline 2 oxoglutarate 4 dioxygenase beta subunit antibody
    • PROHB antibody
    • Prolyl 4 hydroxylase beta polypeptide antibody
    • Prolyl 4 hydroxylase beta subunit antibody
    • Prolyl 4 hydroxylase subunit beta antibody
    • Prolyl 4-hydroxylase subunit beta antibody
    • Protein disulfide isomerase associated 1 antibody
    • Protein disulfide isomerase, family A, member 1 antibody
    • Protein disulfide isomerase/oxidoreductase antibody
    • Protein disulfide-isomerase antibody
    • Protocollagen hydroxylase antibody
    • Thbp antibody
    • Thyroid hormone binding protein p55 antibody
    • Thyroid hormone binding protein p55 cellular antibody
    • V erb a avian erythroblastic leukemia viral oncogene homolog 2 like antibody
    see all

图片

  • Western blot analysis of P4HB (PDIA1) was performed by loading 25 ug of HepG2 (Lane 1) Hela (Lane 2) and NIH-3T3 (Lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4?C overnight. The membrane was probed with ab2792 at 1:1000 overnight at 4?C and washed in TBST. The membrane was then probed with a HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using a ECL Plus Western Blotting Substrate. Results show a band at approx. 57 kDa.

  • Immunocytochemistry/Immunofluorescence analysis of P4HB (PDIA1) (red) in U2OS 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 ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 550 goat anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing P4HB (PDIA1) ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Overlay histogram showing HeLa cells stained with ab2792 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2792, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-P4HB antibody [RL90] (ab2792) at 1/2000 dilution
    Secondary
    Donkey anti mouse IgG2a at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 57 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 seconds

    See Abreview

  • Anti-P4HB antibody [RL90] (ab2792) at 1/1000 dilution + HT1080 whole cell lysate at 20000 cells

    Secondary
    HRP-conjugated sheep anti-mouse IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 57 kDa why is the actual band size different from the predicted?


    Exposure time: 20 seconds


    10% SDS-PAGE.

    Blocked with 5% milk for 1 hour at 22°C.

    Incubated with the primary for 16 hours at 4°C in PBS + 2.5% milk + 0.05% Tween20.

    See Abreview

  • ab2792 staining P4HB (PDIA1) in MDA MB 231 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 10% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/100 in BSA + 0.02% Tween 20) for 1 hour at 16°C. A DyLight® 550-conjugated goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of P4HB (PDIA1) (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 ab2792 (1:75) for at least 1 hour at room temperature and incubated with Dylight 488 goat anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of P4HB (PDIA1) (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 incubated with ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of P4HB (PDIA1) (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 incubated with ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in p19 Cells.

  • Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in NS-1 Cells.

  • Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in HMVEC Cells.

  • Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in A549 Cells.

  • ab2792 positively staining dog MDCK cell ER (red) at 1/200. Staining was carried out in conjunction with goat anti mouse H and L ( Alexa 546) at 1/1000. The nuclei can be seen stained with Hoechst (blue)
    This image is courtesy of an Abreview submitted by Kun Liu on 20 September 2005. We do not have any further information relating to this image.

  • ab2792 positively staining formaldehyde fixed human Hek293 cells (1/200). This Ab was used in conjunction with goat anti mouse Alexa Fluor® 546 (1/1500). The nuclei has been atained with Hoechst.
    This image is courtesy of an Abreview submitted by Kun Liu on 19th September 2005. We do not have any further information relating to this image.

    See Abreview

  • ab2792 staining P4HB (PDIA1) from human HaCaT keratinocyte cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.25 % Triton ×100 and blocking with 2.5% BSA plus 1% goat serum for 1 hour at 4°C was performed. Samples were incubated with primary antibody, diluted 1/100, for 1 hour at 240C. An Alexa Fluor® 488-conjugated goat polyclonal to mouse IgG was used at dilution at 1/1000 as secondary antibody.

    See Abreview

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing P4HB (PDIA1) ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human lung adenocarcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing P4HB (PDIA1) ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Flow cytometry analysis of P4HB (PDIA1) showing positive staining in the membrane and cytoplasm of K562 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of P4HB (PDIA1) showing positive staining in the membrane and cytoplasm of NIH/3T3 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of P4HB (PDIA1) showing positive staining in the membrane and cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

文献

This product has been referenced in:
  • Yamamoto Y & Sakisaka T The peroxisome biogenesis factors posttranslationally target reticulon homology domain-containing proteins to the endoplasmic reticulum membrane. Sci Rep 8:2322 (2018). Read more (PubMed: 29396426) »
  • Lee SM  et al. Recognition of Double-Stranded RNA and Regulation of Interferon Pathway by Toll-Like Receptor 10. Front Immunol 9:516 (2018). Read more (PubMed: 29616030) »
See all 119 Publications for this product

客户评价及客户问答

1-10 of 64 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Cell lysate - other (Macrophages)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
Macrophages
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

提交于 Nov 08 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (heart)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
20 µg
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

提交于 Sep 07 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (heart)
Permeabilization
Yes - 0.2% Triton-X
Specification
heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Sep 03 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Hamster Cell (Chinese Hamster Ovary)
Permeabilization
Yes - 0,1 % TritonX-100 for 5 min @ RT
Specification
Chinese Hamster Ovary
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Dec 20 2016

Application
IHC - Wholemount
Sample
Drosophila melanogaster Tissue (pupae wings)
Specification
pupae wings

Abcam user community

Verified customer

提交于 Apr 23 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MDA MB 231 cells)
Permeabilization
Yes - 1% Triton
Specification
MDA MB 231 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Feb 19 2015

Application
Western blot
Sample
Mouse Cell lysate - whole cell (NIH 3T3)
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Loading amount
50 µg
Specification
NIH 3T3
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

提交于 Jul 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
African green monkey Cell (COS-7)
Permeabilization
Yes - Saponin 0.05%
Specification
COS-7
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Jan 31 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (embryonic ectoderm)
Permeabilization
Yes - tween
Specification
embryonic ectoderm
Blocking step
BSA+milk+serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Dr. Sally Moody

Verified customer

提交于 May 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeoaRG)
Permeabilization
Yes - 0.1% Saponin
Specification
HeoaRG
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Apr 11 2013

1-10 of 64 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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