Anti-P4HB 抗体 [RL90] (ab2792)
Key features and details
- Mouse monoclonal [RL90] to P4HB
- Suitable for: ELISA, Inhibition Assay, IHC-Fr, IHC-P, WB, IP, Flow Cyt, ICC/IF, Electron Microscopy
- Reacts with: Mouse, Rat, Hamster, Dog, Human, Pig, Monkey, African green monkey
- Isotype: IgG2a
製品の概要
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製品名
Anti-P4HB antibody [RL90]
P4HB 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [RL90] to P4HB -
由来種
Mouse -
アプリケーション
適用あり: ELISA, Inhibition Assay, IHC-Fr, IHC-P, WB, IP, Flow Cyt, ICC/IF, Electron Microscopymore details -
種交差性
交差種: Mouse, Rat, Hamster, Dog, Human, Pig, Monkey, African green monkey
交差が予測される動物種: Drosophila melanogaster -
免疫原
Full length native protein (purified) corresponding to Rat P4HB. Purified rat PDIA1/P4HB protein.
Database link: P04785 -
ポジティブ・コントロール
- WB: HeLa, A-431,NIH/3T3, A-375, Hep G2, HL-60, and PC-12.
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特記事項
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
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
RL90 -
アイソタイプ
IgG2a -
研究分野
関連製品
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Assay kits
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab2792の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ELISA |
Use at an assay dependent concentration.
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Inhibition Assay |
Use at an assay dependent concentration.
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IHC-Fr | (1) |
Use at an assay dependent concentration.
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IHC-P | (2) |
1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
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WB | (15) |
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).
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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. |
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Flow Cyt |
Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (26) |
Use at an assay dependent concentration.
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Electron Microscopy |
Use at an assay dependent concentration. PubMed: 21886772
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特記事項 |
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ELISA
Use at an assay dependent concentration. |
Inhibition Assay
Use at an assay dependent concentration. |
IHC-Fr
Use at an assay dependent concentration. |
IHC-P
1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol. |
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). |
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. |
Flow Cyt
Use 0.5µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use at an assay dependent concentration. |
Electron Microscopy
Use at an assay dependent concentration. PubMed: 21886772 |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the protein disulfide isomerase family.
Contains 2 thioredoxin domains. -
細胞内局在
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
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参照データベース
- 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
see all -
別名
- Cellular thyroid hormone binding protein antibody
- Cellular thyroid hormone-binding protein antibody
- Collagen prolyl 4 hydroxylase beta antibody
see all
画像
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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 kDa why is the actual band size different from the predicted? -
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.
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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.
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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.
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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.
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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
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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 seconds10% 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.
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Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)This image is courtesy of an anonymous 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.
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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.
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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.
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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.
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Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in p19 Cells.
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Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in NS-1 Cells.
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Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in HMVEC Cells.
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Immunocytochemistry/Immunofluorescent analysis of P4HB (PDIA1) using ab2792 shows staining in A549 Cells.
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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. -
Immunocytochemistry/ Immunofluorescence - Anti-P4HB antibody [RL90] (ab2792)This image is a courtesy of Anonymous 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.
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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.
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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.
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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.
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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.
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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (195)
ab2792 は 195 報の論文で使用されています。
- Lee DS & Kim JE Protein disulfide isomerase-mediated S-nitrosylation facilitates surface expression of P2X7 receptor following status epilepticus. J Neuroinflammation 18:14 (2021). PubMed: 33407649
- Jiang S et al. Hydrogen sulphide reduces hyperhomocysteinaemia-induced endothelial ER stress by sulfhydrating protein disulphide isomerase to attenuate atherosclerosis. J Cell Mol Med 25:3437-3448 (2021). PubMed: 33675119
- Petkovic M et al. TMEM16K is an interorganelle regulator of endosomal sorting. Nat Commun 11:3298 (2020). PubMed: 32620747
- Tischer A et al. Evidence for the Misfolding of the A1 Domain within Multimeric von Willebrand Factor in Type 2 von Willebrand Disease. J Mol Biol 432:305-323 (2020). PubMed: 31628947
- Jia J et al. AMPK, a Regulator of Metabolism and Autophagy, Is Activated by Lysosomal Damage via a Novel Galectin-Directed Ubiquitin Signal Transduction System. Mol Cell 77:951-969.e9 (2020). PubMed: 31995728