Anti-P4HB antibody [RL90]

• 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: ELISA, Inhibition Assay, IHC-Fr, IHC-P, WB, IP, Flow Cyt, ICC/IF, Electron Microscopymore 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

  Full length native protein (purified) corresponding to Rat P4HB. Purified rat PDIA1/P4HB protein.
  Database link: P04785

• 阳性对照

• WB: HeLa, A-431,NIH/3T3, A-375, Hep G2, HL-60, and PC-12. ICC/IF: HeLa, A-431, MCF7, A549 and U2OS cells

• 常规说明

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  Preservative: 0.05% Sodium azide
  Constituents: 0.1% BSA, PBS

• 浓度

• 100 µl 浓度为 1 mg/ml

• 纯度

  Protein A purified

• 克隆

  单克隆

• 克隆编号

  RL90

• 同种型

  IgG2a

The Abpromise guarantee

Abpromise™承诺保证使用ab2792于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

数据库链接

• Entrez Gene: 483369 Dog

• Entrez Gene: 39651 Drosophila melanogaster

• Entrez Gene: 5034 Human

• Entrez Gene: 18453 Mouse

• Entrez Gene: 110255932 Pig

• Entrez Gene: 25506 Rat

• Omim: 176790 Human

• SwissProt: P54399 Drosophila melanogaster

• SwissProt: P07237 Human

• SwissProt: P09103 Mouse

• SwissProt: P04785 Rat

• Unigene: 2710 Drosophila melanogaster

• Unigene: 464336 Human

• Unigene: 708779 Human

• Unigene: 16660 Mouse

• Unigene: 392196 Mouse

• Unigene: 4234 Rat

别名

• Cellular thyroid hormone binding protein antibody

• Cellular thyroid hormone-binding protein antibody

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• 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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  ab2792 staining P4HB in wild-type A431 cells (top panel) and P4HB knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2792 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 4% paraformaldehyde (10 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Western blot - Anti-P4HB antibody [RL90] (ab2792)

  All lanes : Anti-P4HB antibody [RL90] (ab2792) at 1/2000 dilution
  
  Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
  Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell lysate
  Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
  Lane 4 : A-375 (human malignant melanoma cell line) whole cell lysate
  Lane 5 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
  Lane 6 : HL-60 (human promyelocytic leukemia cell line) whole cell lysate
  Lane 7 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
  
  Lysates/proteins at 30 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse IgG H+L (HRP) at 1/4000 dilution
  
  Predicted band size: 58 kDa
  Observed band size: 57 kDawhy is the actual band size different from the predicted?
  
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  ab2792 staining P4HB in MCF7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2792 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 4% paraformaldehyde (10 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  ab2792 staining P4HB in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2792 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  ab2792 staining P4HB in A549 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2792 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  ab2792 staining P4HB in U2OS cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2792 at 0.2µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150084, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 4% paraformaldehyde (10 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P4HB antibody [RL90] (ab2792)

  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.
  
  

• 

  Flow Cytometry - Anti-P4HB antibody [RL90] (ab2792)

  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.
  
  

• 

  Western blot - Anti-P4HB antibody [RL90] (ab2792)

  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.

• 

  Western blot - Anti-P4HB antibody [RL90] (ab2792)This image is courtesy of an anonymous Abreview

  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 kDawhy is the actual band size different from the predicted?
  
  
  Exposure time: 20 seconds
  

  See Abreview

• 

  Western blot - Anti-P4HB antibody [RL90] (ab2792)

  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 kDawhy 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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)

  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.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P4HB antibody [RL90] (ab2792)

  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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P4HB antibody [RL90] (ab2792)

  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 - Anti-P4HB antibody [RL90] (ab2792)

  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 - Anti-P4HB antibody [RL90] (ab2792)

  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 - Anti-P4HB antibody [RL90] (ab2792)

  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.