GRP78/HSPA5 Antibody - BSA Free

Simple Western: GRP78/HSPA5 Antibody [NBP1-06274] - Image shows a specific band for GRP78 in 0.1 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - C. elegans granulins impair organismal fitness and resistance to ER stress. Total worm lysates from synchronized day 1 adult granulin-expressing animals were immunoblotted with an anti-GRP78/HSPA5 antibody (3 biological replicates). Anti-actin was used as a loading control. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pgen.1008295), licensed under a CC-BY licence.

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - Analysis of MAM-associated protein levels in control and patients' fibroblasts.(A) Equal amounts from one control and the five patients were loaded (50 ug of total cell lysates) and subjected to Western Blot with anti-Grp75, anti-sigma-1R and anti-Mfn2 antibodies. We used anti-GAPDH antibody to ensure equal amounts of protein loaded in each lane. This result is representative of three independent experiments. PLoS One. 2016 Mar 9;11(3):e0150357. doi: 10.1371/journal.pone.0150357

Immunocytochemistry/Immunofluorescence: GRP78/HSPA5 Antibody [NBP1-06274] - NIH-3T3 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-GRP78/HSPA5 at 2 ug/mL overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - Total protein from human HeLa and A431 cells, mouse 3T3 cells and rat PC12 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/mL anti-GPR78 in blocking buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunocytochemistry/Immunofluorescence: GRP78/HSPA5 Antibody [NBP1-06274] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-GRP78/HSPA5 at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on NIH3T3 cells with GRP78/HSPA5 Antibody NBP1-06274AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - Detection of Bip/Grp78 on HeLa whole cell extracts using NBP1-06274.

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - Analysis using Alexa Fluor (R) 488 conjugate of NBP1-06274. An intracellular stain was performed on HeLa cells with GPR78/HSPA5 antibody NBP1-06274 (blue) and a matched isotype control NBP2-24893 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. 1 ug of antibody was added to 100 uL of staining buffer and cells were incubated for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on Jurkat cells with Alexa Fluor 647 conjugate of GPR78/HSPA5 antibody NBP1-06274AF647 (blue) and a matched isotype control NBP2-24893AF647 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on HeLa with NBP1-06274 and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550.

• Reactivity: Hu, Mu, Rt, Ce, Ch, Dr, Sh, Ze, Bv, Pm
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IHC-FrFl
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

GRP78/HSPA5 Antibody - BSA Free Summary

• Immunogen: Synthetic peptide made to an internal portion of human GRP78 (within residues 250-300). [Swiss-Prot# P11021]
• Localization: Endoplasmic reticulum lumen. Melanosome. Note: Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
• Marker: ER Stress Marker
• Predicted Species: Bovine (100%), Primate (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HSPA5
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:150
  • Immunocytochemistry/Immunofluorescence 1:50
  • Immunohistochemistry Free-Floating
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Simple Western 1:25
  • Western Blot 0.5 ug/ml
• Application Notes: In Western blot a band is seen at ~78 kDa. For information on the flow cytometry application, see the publication PMID: 20208072. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 24089213). Use in Immunohistochemistry-paraffin reported in scientific literature (PMID: 24089213). Use in Immunohistochemistry free floating reported in scientific literature (PMID: 26329458).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 78 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  HeLa Whole Cell Lysate

• Control Peptide:

  GRP78/HSPA5 Peptide (NBP1-06274PEP)

Reactivity Notes

Use in Zebrafish reported in scientific literature (PMID:34327238) C. elegans reactivity reported in scientific literature (PMID: 31398187).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for GRP78/HSPA5 Antibody - BSA Free

• 78 kDa glucose-regulated protein
• BIP
• Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78
• FLJ26106
• GRP78
• GRP-78
• GRP78BIP
• Heat shock 70 kDa protein 5
• heat shock 70kD protein 5 (glucose-regulated protein, 78kD)
• heat shock 70kDa protein 5 (glucose-regulated protein, 78kDa)
• HSP70-5
• HSPA5
• Immunoglobulin heavy chain-binding protein
• MIF2

Background

HSP70 family member GRP78 (78 kD glucose-regulated protein) is localized in ER lumen as well as melanosomes (stage I to stage IV) where it facilitates protein folding/assembly, protein quality control, Ca2+ binding, and regulates ER stress signaling. GRP78 interacts with DNAJC1 and is a component of EIF2 complex (CELF1/CUGBP1, CALR, CALR3, EIF2S1, EIF2S2, HSP90B1 and HSPA5) as well as chaperone multiprotein complex (DNAJB11, HSP90B1, HSPA5, HYOU, PDIA2, PDIA4, PDIA6, PPIB, SDF2L1, UGT1A1, ERP29). GRP78 also interacts with TMEM132A as well as TRIM21, and may form a complex with ERLEC1, OS9, SEL1L and SYVN1. In cancerous cells, GRP78 overexpression is associated with UPR activation, including upregulation of associated proteins such as PERK, ATF6, CHOP etc that further regulate JNK and NF-kB untimately leading to survival, tumor progression, angiogenesis, resistance to therapy and metastasis. GRP78 is a signaling receptor for activated alpha2-macroglobulin, plasminogen kringle 5, and microplasminogen, and plays role in viral entry of coxsackie B as well as dengue fever viruses. GRP78 is also involved in regulation of tissue factor procoagulant activity and functions as a receptor for angiogenic peptides via VEGFRs independent mechanism. Cell surface GRP78 is found associated with diverse proteins including VDAC, MHC-I, Cripto and DnaJ-like protein MTJ-1. Because GRP78 is present on the surface of cancer cells but not on normal cells in vivo, it provide an exciting opportunity for cancer targeting.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GRP78/HSPA5 Antibody (NBP1-06274)(38)

Publications using NBP1-06274

• Shrestha S, Erikson G, Lyon J et al. Aging compromises human islet beta cell function and identity by decreasing transcription factor activity and inducing ER stress Science advances 2022-10-07 [PMID: 36197983] (IHC-P, Human)
• Lemoine H, Raud L, Foulquier F et al. Monoallelic pathogenic ALG5 variants cause atypical polycystic kidney disease and interstitial fibrosis American journal of human genetics 2022-07-20 [PMID: 35896117]
• Aggarwal P, Liu Z, Cheng GQ et al. Disruption of the lipolysis pathway results in stem cell death through a sterile immunity-like pathway in adult Drosophila Cell reports 2022-06-21 [PMID: 35732115] (IF/IHC, Drosophilia)
• Li Q, Fan X, Lu W et al. Novel NPR2 Gene Mutations Affect Chondrocytes Function via ER Stress in Short Stature Cells 2022-04-08 [PMID: 35455946] (WB)
• Yuan X, Shen Q, Ma W Long Noncoding RNA Hotair Promotes the Progression and Immune Escape in Laryngeal Squamous Cell Carcinoma through MicroRNA-30a/GRP78/PD-L1 Axis Journal of immunology research 2022-04-04 [PMID: 35419461] (WB, Human)
• Chou CW, Yang RY, Chan LC et al. The stabilization of PD-L1 by the endoplasmic reticulum stress protein GRP78 in triple-negative breast cancer American journal of cancer research 2020-08-01 [PMID: 32905506] (WB)
• Shaiken T, Grimm S, Siam M et al. Transcriptome, Proteome, and Protein Synthesis Within the Intracellular Cytomatrix SSRN Electronic Journal 2022-03-31
• Chen X, Li Z, Lv y et al. Comparative proteomic identification of capacitated and non-capacitated sperm of yanbian yellow Cattle Theriogenology 2022-03-01 [PMID: 35421774] (WB, Bovine)
• Rossin F, Avitabile E, Catarinella G et al. Reticulon-1C Involvement in Muscle Regeneration Metabolites 2021-12-08 [PMID: 34940613]
• Stojkovska I, Wani WY, Zunke F Et al. Rescue of alpha-synuclein aggregation in Parkinson's patient neurons by synergistic enhancement of ER proteostasis and protein trafficking Neuron 2021-11-09 [PMID: 34793693]
• Arquier N, Bjordal M, Hammann P et al. Brain adiponectin signaling controls peripheral insulin response in Drosophila Nature communications 2021-09-24 [PMID: 34561451] (WB, Drosophila)
• VerPlank JJS, Gawron J, Silvestri NJ Et al. Raising cGMP restores proteasome function and myelination in mice with a proteotoxic neuropathy Brain : a journal of neurology 2021-08-11 [PMID: 34382059] (WB)
• Li Lp, Zhong J, Li Mh Et Al. Disruption of MAP7D1 Gene Function Increases the Risk of Doxorubicin-Induced Cardiomyopathy and Heart Failure BioMed research international 2021-07-15 [PMID: 34327238] (WB)
• Nam YW, Kong D, Wang D, et al. Differential modulation of SK channel subtypes by phosphorylation Cell calcium 2021-01-06 [PMID: 33422768] (WB, Mouse)
• Arioka Y, Shishido E, Kushima I et al. Chromosome 22q11.2 deletion causes PERK-dependent vulnerability in dopaminergic neurons EBioMedicine 2020-12-17 [PMID: 33341442] (WB, Human)
• Balcioglu O, Heinz RE, Freeman DW et al. CRIPTO antagonist ALK4L75A-Fc inhibits breast cancer cell plasticity and adaptation to stress Breast Cancer Res 2020-11-13 [PMID: 33187540] (FLOW)
• Yousuf MS, Samtleben S, Lamothe SM et al. Endoplasmic reticulum stress in the dorsal root ganglia regulates large-conductance potassium channels and contributes to pain in a model of multiple sclerosis FASEB J. 2020-07-17 [PMID: 32677089] (WB, IF/IHC, Human)
• Scapin C, Ferri C, Pettinato E et al. Enhanced axonal Neuregulin-1 type-III signaling ameliorates disease severity in a CMT1B mouse model bioRxiv 2018-08-09 (WB, Mouse)
• Asahina, M, Fujinawa, R Et al. Ngly1 -/- rats develop neurodegenerative phenotypes and pathological abnormalities in their peripheral and central nervous systems. Hum Mol Genet 2020-06-27 [PMID: 32259258] (FLOW, Human)
• Butler VJ, Gao F, Corrales CI et al. Age- and stress-associated C. elegans granulins impair lysosomal function and induce a compensatory HLH-30/TFEB transcriptional response PLoS Genet. 2019-08-01 [PMID: 31398187] (WB, C. elegans)
• Subramanian A, Capalbo A, Iyengar NR et al. Auto-regulation of Secretory Flux by Sensing and Responding to the Folded Cargo Protein Load in the Endoplasmic Reticulum Cell 2019-03-07 [PMID: 30849374] (WB)
• Wang Y, Wang X, Li H et al. Binge ethanol exposure induces endoplasmic reticulum stress in the brain of adult mice Toxicol. Appl. Pharmacol. 2018-08-13 [PMID: 30114398] (WB, Mouse)
• Ren Z, Wang X, Yang F et al. Ethanol-induced Damage to the Developing Spinal Cord: The Involvement of CCR2 Signaling Biochim. Biophys. Acta 2017-08-01 [PMID: 28778590] (WB, Mouse)
• Liu YQ, Jia MQ, Xie ZH et al. Arrestins contribute to amyloid beta-induced cell death via modulation of autophagy and the alpha7nAch receptor in SH-SY5Y cells. Sci Rep. 2017-06-13 [PMID: 28611418] (WB, Human)
• Liu Hua, Wang Zhe, Nowicki Michael J. Caspase-12 mediates carbon tetrachloride-induced hepatocyte apoptosis in mice. World J Gastroenterol. 2014-12-28 [PMID: 25561786] (WB, Mouse)
• Adolph TE, Tomczak MF, Niederreiter L et al. Paneth cells as a site of origin for intestinal inflammation. Nature. [PMID: 24089213] (ICC/IF, IHC-P, Mouse)
• Zhang Y, Liu R, Ni M et al. Cell surface relocalization of the endoplasmic reticulum chaperone and unfolded protein response regulator GRP78/BiP. J Biol Chem. 2010-05-14 [PMID: 20208072]
• Butler V, Cortopassi W, Argouarch A et al. C. elegans granulins promote an age-associated decline in protein homeostasis via lysosomal protease inhibition. CST. 2018-11-12 (WB)
• Kunde DA, Chong WC, Nerurkar PV et al. Bitter melon protects against ER stress in LS174T colonic epithelial cells. BMC Complement Altern Med 2017-01-03 [PMID: 28049460] (WB)
• Sidoli M, Musner N, Silvestri N et al. Ablation of Perk in Schwann Cells Improves Myelination in the S63del Charcot-Marie-Tooth 1B Mouse. J. Neurosci. 2016-11-02 [PMID: 27807175] (WB, Mouse)
• Ren Z, Yang F, Wang X et al. Chronic plus binge ethanol exposure causes more severe pancreatic injury and inflammation. Toxicol. Appl. Pharmacol. 2016-08-15 [PMID: 27538709] (WB, Mouse)
• Ren Z, Wang X, Xu M et al. Binge ethanol exposure causes endoplasmic reticulum stress, oxidative stress and tissue injury in the pancreas. Oncotarget. 2016-08-23 [PMID: 27527870] (ICC/IF, Rat)
• Niccoli T, Cabecinha M, Tillmann A et al. Increased Glucose Transport into Neurons Rescues Abeta Toxicity in Drosophila. Curr. Biol. 2016-08-09 [PMID: 27524482] (WB, Drosophila)
• Rasche L, Menoret E, Dubljevic V et al. A GRP78-directed monoclonal antibody recaptures response in refractory multiple myeloma with extramedullary involvement. Clin. Cancer Res. 2016-03-30 [PMID: 27029491] (FLOW, Human)
  
  Details:
  This publication used the DL488 conjugated form of this antibody (NBP1-06274G)
• Martinez-Pizarro A, Desviat LR, Ugarte M et al. Endoplasmic Reticulum Stress and Autophagy in Homocystinuria Patients with Remethylation Defects. PLoS ONE. 2016-03-10 [PMID: 26959487] (WB, Human)
• Henry FE, Sugino K, Tozer A et al. Cell type-specific transcriptomics of hypothalamic energy-sensing neuron responses to weight-loss. Elife 2015-09-02 [PMID: 26329458] (IHC-FrFl, Mouse)
  
  Details:
  GRP78 antibody was used at 1:4000 dilution for IHC- free floating sections on brains of Agrp(Cre), Agrptm1(cre)Lowl/J), Ai9 (ROSA-loxPStoploxP-tdTomato), Pomc(topazFP), and Npy(hrGFP) mice.
• Grevengoed TJ, Martin SA, Katunga L et al. Acyl-CoA synthetase 1 deficiency alters cardiolipin species and impairs mitochondrial function J. Lipid Res. 2015-08-01 [PMID: 26136511] (ICC/IF)
• Pepping JK, Freeman LG, Gupta S et al. NOX2 Deficiency Attenuates Markers of Adiposopathy and Brain Injury Induced by High Fat Diet Am J Physiol Endocrinol Metab 2012-12-11 [PMID: 23233541] (WB, Mouse)

Reviews for GRP78/HSPA5 Antibody (NBP1-06274) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Other.

reviewed by: Muhammad Yousuf WB Mouse 03/23/2019

Summary

• Application: Western Blot
• Sample Tested: Mouse dorsal root ganglia
• Species: Mouse
• Lot: G-1

Comments

• Comments: Primary antibody BiP incubated at 1:1000 at 4 degrees overnight. Secondary antibody goat anti-rabbit HRP incubated at 1:10,000 for 1 hour at room temperature. Visualized using ECL.

reviewed by: Yonghe Ding WB Other 07/27/2016

Summary

• Application: Western Blot
• Sample Tested: Whole embryos lysate
• Species: Other

FAQs for GRP78/HSPA5 Antibody (NBP1-06274). (Showing 1 - 1 of 1 FAQs).

1. I have received the Novus Cat # for a human GRP78-specific rabbit Ab from a lab that had used it and achieved some nice results with it. They suggest we use the same Ab to reproduce their data. However, I have not been able to find a product with that Cat # in your online search engine. Perhaps the product has been discontinued, or was given a different Cat #? Could you please look into this? The Ab they used was: GRP78 Cat # NBP1-49339, Lot # A113911. In case the product is no longer available, could you please recommend something that should be equivalent or very similar to it?
   • NBP1-49339 has been discontinued but NBP1-06274 uses a similar antigen that falls between amino acids 250 and 300 and works in Western blot and ICC. ICC and flow use essentially the same techniques, so it should work just fine if you were using it in flow or Western blot before.

Bioinformatics

• Gene Symbol: HSPA5