概述

  • 产品名称Anti-GRP78 BiP抗体
    参阅全部 GRP78 BiP 一抗
  • 描述
    兔多克隆抗体to GRP78 BiP
  • 经测试应用适用于: IHC-FoFr, IP, ICC/IF, WB, Electron Microscopy, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Dog, Human, Pig, African Green Monkey, Chinese Hamster
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Mouse GRP78 BiP.

    (Peptide available as ab22410.)

  • 阳性对照
    • This antibody gave a positive signal in both Mouse and Rat liver tissue lysates as well as the following whole cell lysates: CHO-K1; HeLa.

性能

  • 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: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab21685 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
IHC-FoFr Use at an assay dependent concentration.
IP Use a concentration of 2 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).Can be blocked with Mouse GRP78 BiP peptide (ab22410).
Electron Microscopy 1/250.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能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.
  • 疾病相关Autoantigen in rheumatoid arthritis.
  • 序列相似性Belongs to the heat shock protein 70 family.
  • 细胞定位Endoplasmic reticulum lumen. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 78 kDa glucose regulated protein antibody
    • 78 kDa glucose-regulated protein antibody
    • AL022860 antibody
    • AU019543 antibody
    • BIP antibody
    • D2Wsu141e antibody
    • D2Wsu17e antibody
    • Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 antibody
    • Endoplasmic reticulum lumenal Ca2+ binding protein grp78 antibody
    • Epididymis secretory sperm binding protein Li 89n antibody
    • FLJ26106 antibody
    • Glucose Regulated Protein 78kDa antibody
    • GRP 78 antibody
    • GRP-78 antibody
    • GRP78 antibody
    • GRP78_HUMAN antibody
    • Heat shock 70 kDa protein 5 antibody
    • Heat Shock 70kDa Protein 5 antibody
    • Heat shock protein family A (Hsp70) member 5 antibody
    • HEL S 89n antibody
    • Hsce70 antibody
    • HSPA 5 antibody
    • HSPA5 antibody
    • Immunoglobulin Heavy Chain Binding Protein antibody
    • Immunoglobulin heavy chain-binding protein antibody
    • mBiP antibody
    • MIF2 antibody
    • Sez7 antibody
    see all

Anti-GRP78 BiP antibody 图像

  • ICC/IF image of ab21685 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab21685, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • 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.
  • All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml

    Lane 1 : CHO-K1 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 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
    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.

    See Abreview

  • ICC/IF image of ab21685 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21685, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ICC/IF image of ab21685 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21685, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml

    Lane 1 : CHO-K1 cell lysate 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
    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
  • ab21685 staining GRP78 BiP in African Green Monkey COS-7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 23°C. Samples were incubated with primary antibody (1/1000 in PBS-BSA) for 1 hour at 23°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • ab21685 at a 1/500 dilution staining GRP78 BiP in mouse retinal pigment epithelium primary cells by Immunocytochemistry/ Immunofluorescence, incubated for 16 hours at 4°C. PFA fixed. Blocked with 5% serum for 20 minutes at 25°C. Secondary used at a 1/500 dilution polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488 (green). Nuclei were counterstained with DAPI (blue).

    See Abreview

  • ab21685 at a 1/500 dilution staining GRP78 BiP in Dog MDCK II cells by Immunocytochemistry/ Immunofluorescence, incubated for 16 hours at 4°C. PFA fixed. Blocked with 5% serum for 20 minutes at 25°C. Secondary used at a 1/500 dilution polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488 (green). Nuclei were counterstained with DAPI (blue).

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 78 kDa

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 78 kDa


    Exposure time : 10 seconds

Anti-GRP78 BiP antibody (ab21685)参考文献

This product has been referenced in:
  • Gai Z  et al. Farnesoid X Receptor Protects against Kidney Injury in Uninephrectomized Obese Mice. J Biol Chem 291:2397-411 (2016). Read more (PubMed: 26655953) »
  • Kang K  et al. Carnosic acid slows photoreceptor degeneration in the Pde6b(rd10) mouse model of retinitis pigmentosa. Sci Rep 6:22632 (2016). IHC, WB . Read more (PubMed: 26961159) »

See all 58 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Rat hepatocytes)
Permeabilization Yes - 0.2% Triton X-100
Specification Rat hepatocytes
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

提交于 Nov 28 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Loading amount 25 µg
Specification Rat hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Nov 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (WIF-B cells, the hepatoma-derived hybrid cell line)
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Loading amount 30 µg
Specification WIF-B cells, the hepatoma-derived hybrid cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Nov 22 2016

Application Western blot
Sample Human Cell lysate - whole cell (MDA435 Cells)
Gel Running Conditions Reduced Denaturing (4-20% Tris Glycin Gel)
Loading amount 50000 cells
Specification MDA435 Cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Sep 13 2016

Application Western blot
Sample Sheep Cell lysate - whole cell (undifferentiated ovine preadipocytes)
Gel Running Conditions Reduced Denaturing (10% PAG)
Loading amount 10 µg
Specification undifferentiated ovine preadipocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Mr. Jeremy Gingrich

Verified customer

提交于 Sep 02 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (B16 cell line)
Permeabilization Yes - 0.25% Triton-X100 in PBS
Specification B16 cell line
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 12 2016

Application Western blot
Sample Human Cell lysate - whole cell (vascular smooth muscle)
Gel Running Conditions Non-reduced Denaturing (8%)
Loading amount 10 µg
Specification vascular smooth muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

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提交于 Feb 02 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Mouse Embryonic Fibroblast (MEF))
Permeabilization Yes - 0.5% TX-100
Specification Mouse Embryonic Fibroblast (MEF)
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Ms. Samantha O'Hara

Verified customer

提交于 Oct 27 2015

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (retina)
Specification retina
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 09 2015

Application Western blot
Loading amount 80 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (gel 7.5%)
Sample Mouse Tissue lysate - whole (retina)
Specification retina
Treatment 5% Milk for 1h RT
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Minzhong Yu

Verified customer

提交于 Feb 09 2015

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