Anti-GRP78 BiP 抗体 (ab21685)
Key features and details
- Rabbit polyclonal to GRP78 BiP
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Human, Chinese hamster, Recombinant fragment
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-GRP78 BiP antibody
GRP78 BiP 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to GRP78 BiP -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human, Chinese hamster, Recombinant fragment
交差が予測される動物種: Dog, Pig, African green monkey
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免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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特記事項
Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls.
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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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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ab21685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 |
|---|---|---|
| ICC/IF | (25) |
Use a concentration of 1 - 5 µg/ml.
We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody. |
| WB | (31) |
Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).
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| IHC-P | (4) |
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| 特記事項 |
|---|
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ICC/IF
Use a concentration of 1 - 5 µg/ml. We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa). |
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IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ターゲット情報
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機能
Probably plays a role in facilitating the assembly of multimeric protein complexes inside the endoplasmic reticulum. Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10, probably to facilitate the release of DNAJC10 from its substrate. -
関連疾患
Autoantigen in rheumatoid arthritis. -
配列類似性
Belongs to the heat shock protein 70 family. -
細胞内局在
Endoplasmic reticulum lumen. Melanosome. Cytoplasm. 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: 100689305 Chinese hamster
- Entrez Gene: 480726 Dog
- Entrez Gene: 3309 Human
- Entrez Gene: 14828 Mouse
- Entrez Gene: 407060 Pig
- Entrez Gene: 25617 Rat
- Omim: 138120 Human
- SwissProt: P11021 Human
see all -
別名
- 78 kDa glucose regulated protein antibody
- 78 kDa glucose-regulated protein antibody
- AL022860 antibody
see all
画像
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Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa treated with 2.5 µg/ml tunicamycin for 24h whole cell lysates
Lane 3 : HUVEC (Human umbilical vein endothelial cell) whole cell lysates
Lane 4 : HUEVC (Human umbilical vein endothelial cell) treated with 10 µg/ml tunicamycin for 48h whole cell lysates
Lane 5 : Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 6 : Raw 264.7 treated with 5 µg/ml tunicamycin for 18h whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 78 kDa
Observed band size: 78 kDa
Exposure time: 3 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)Image from Wang Y et al., PLoS One. 2017;12(7):e0180731. Fig 3.; doi: 10.1371/journal.pone.0180731. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/HEK-293 cells were cultured on coverslips and transiently transfected with wild-type murine IGSF1-2-Myc/His, premeabilized, and processed for double-label immunofluorescence with the Myc antibody (green) and an antibody against GRP78 BiP (red). The overlay is shown in yellow. Nuclei were stained with DAPI. Images were captured by confocal microscopy. Scale bar, 10 μm.
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)Image from Friebe S et al., PLoS One. 2015;10(3):e0119864. Fig 3.; doi: 10.1371/journal.pone.0119864. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Localization of TEM8 and CMG2 glycosylation mutants.
A) Immunofluorescence of transiently transfected HeLa cells. Cells were transfected for 48h with the respective cDNAs. Cells were fixed, permeabilized and stained for TEM8-HA, endogenous BiP and Hoechst. Scalebars represent 10 μm.
B) Immunofluorescence of transiently transfected HeLa cells. Cells were transfected for 48h with the respective cDNAs. Cells were fixed, permeabilized and stained for CMG2-V5, endogenous BiP and Hoechst. Scalebars represent 10 μm.
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Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : CHO-K1 (chinese hamster ovary cell line) whole cell lysate at 20 µg
Lane 2 : Liver (Mouse) Tissue Lysate at 20 µg
Lane 3 : Rat liver whole cell lysate at 20 µg
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 5 : CHO-K1 whole cell lysate at 20 µg/ml with Mouse GRP78 BiP peptide (ab22410) at 1 µg
Lane 6 : Liver (Mouse) Tissue Lysate at 20 µg with Mouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 7 : Rat liver whole cell lysate at 20 µg with Mouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 8 : HeLa whole cell lysate at 20 µg with Mouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Secondary
All lanes : Goat anti Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.ab21685 recognises a band of ~ 75 kDa in CHO, mouse liver, rat liver and HeLa whole cell lysates, corresponding to GRP78 BiP. This band is quenched by the addition of the immunizing peptide, ab22410.
ab21685 also detects a 100 kDa band in Western Blot. We are unsure of the identity of this protein.
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)ab21685 stained in Hela cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab21685 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (pseudo-colored red) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab21685)IHC image of GRP78 BiP staining in human liver carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab21685, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : CHO-K1 (chinese hamster ovary cell line) Whole Cell Lysate
Lane 2 : Liver (Mouse) Tissue Lysate at 10 µg
Lane 3 : Liver (Rat) Tissue Lysate at 10 µg
Lane 4 : HeLa (human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Lanes 1 & 4 : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Lanes 2-3 : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 78 kDa
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 :Recombinant human GRP78 BiP protein (Active) (ab78432) at 0.1 µg
Lane 2 :Recombinant human GRP78 BiP protein (Active) (ab78432) at 0.01 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 78 kDa
Exposure time: 10 seconds -
Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)This image is courtesy of an anonymous abreview.Immunocytochemistry/ Immunofluorescence analysis of MDA-MB-435S tumor cell line cells labeling GRP78 BiP with ab21685 at 1/400 dilution. Cells were fixed in formaldehyde and permeabilized with 0.25% Triton-X100 in PBS for 10 minutes. Blocking was done with 1%BSA for 1 hour at 20°C; followed by staining with ab21685 at 1/400 for 18 hours. Undiluted ab150077, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody was used. DAPI was used to counterstain.
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 (463)
ab21685 は 463 報の論文で使用されています。
- Gao LJ et al. Ligustilide alleviates neurotoxicity in SH-SY5Y cells induced by Aß25-35 via regulating endoplasmic reticulum stress and autophagy. Phytother Res 35:1572-1584 (2021). PubMed: 33111362
- Zhang Y et al. Duck enteritis virus infection suppresses viability and induces apoptosis and endoplasmic reticulum stress in duck embryo fibroblast cells via the regulation of Ca2. J Vet Med Sci 83:549-557 (2021). PubMed: 33116004
- Jin Y et al. Depletion of Adipocyte Becn1 Leads to Lipodystrophy and Metabolic Dysregulation. Diabetes 70:182-195 (2021). PubMed: 33046512
- Li J et al. Overexpression of lncRNA Dancr inhibits apoptosis and enhances autophagy to protect cardiomyocytes from endoplasmic reticulum stress injury via sponging microRNA-6324. Mol Med Rep 23:N/A (2021). PubMed: 33300079
- Li KK et al. U three protein 14a (UTP14A) promotes tumour proliferation and metastasis via the PERK/eIF2a/GRP78 signalling pathway in oesophageal squamous cell carcinoma. J Cancer 12:134-140 (2021). PubMed: 33391409