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GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751978

GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC(Intra)

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Green monkey

  • Pig

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

GRP94 Recombinant Rabbit Monoclonal Antibody [PSH25-59] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Species Reactivity

Human, Mouse, Rat, Green monkey, Pig

Validated Applications

WB, IF-Cell, IHC-P, FC(Intra), IP

Target Molecular Weight

Predicted band size: 92 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cell) cell lysate, MCF7 (Human breast cancer cell) cell lysate, PANC-1 (Human epidermoid carcinoma pancreatic ductal cell) cell lysate, MEF (Mouse embryonic fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, COS-1 (African green monkey kidney fibroblast) cell lysate, Pig brain tissue lysate, Mouse brain tissue lysate, Mouse stomach tissue lysate, Rat brain tissue lysate, Rat stomach tissue lysate.

Conjugation

unconjugated

Clone Number

PSH25-59

Product Features

Form

Liquid

Concentration

0.5 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:10,000

  • IF-Cell

  • 1:500-1:2,500

  • IHC-P

  • 1:5,000

  • FC(Intra)

  • 1 μg/mL

  • IP

  • 1-2μg/sample

Target

Function

Heat shock protein 90kDa beta member 1 (HSP90B1), known also as endoplasmin, gp96, grp94, or ERp99, is a chaperone protein that in humans is encoded by the HSP90B1 gene. HSP90B1 is an HSP90 paralogue that is found in the endoplasmic reticulum. It plays critical roles in folding proteins in the secretory pathway such as Toll-like receptors and integrins. It has been implicated as an essential immune chaperone to regulate both innate and adaptive immunity. Tumor-derived HSP90B1 (vitespen) has entered clinical trials for cancer immunotherapy. GRP94 has been shown to be a target for treatment of a plethora of diseases such as glaucoma, multiple myeloma, and metastatic cancer. grp94 includes 5 distinct amino acids in its primary sequence which creates 2 unique sub-pockets, S1 and S2. These sub-pockets have been utilized in current research in order to inhibit the chaperone since its client proteins seem to be up-regulated in cancer cells.

Background References

1. Xu H et al. Conformationally Restricted Grp94-Selective Inhibitors. ACS Omega. 2025 Jan

2. Ding S et al. AI662270/GRP94 axis couples the unfolded protein response to mitochondrial dynamics during acute myocardial infarction. JCI Insight. 2025 Oct

Subcellular Location

Endoplasmic reticulum lumen, Sarcoplasmic reticulum lumen, Melanosome.

UNIPROT

SwissProt: P14625 Human

SwissProt: P08113 Mouse

SwissProt: Q66HD0 Rat

Synonyms

94 kDa glucose regulated protein antibody

94 kDa glucose-regulated protein antibody

ECGP antibody

Endoplasmin antibody

Endothelial cell (HBMEC) glycoprotein antibody

ENPL_HUMAN antibody

Glucose regulated protein 94kDa antibody

gp96 antibody

gp96 homolog antibody

GRP 94 antibody

Expand

Images

  • Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (<a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>) at 1/10,000 dilution. <br /> <br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate <br />Lane 2: MCF7 (Human breast cancer cell) cell lysate <br />Lane 3: PANC-1 (Human epidermoid carcinoma pancreatic ductal cell) cell lysate <br />Lane 4: MEF (Mouse embryonic fibroblast) cell lysate <br />Lane 5: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate <br />Lane 6: COS-1 (African green monkey kidney fibroblast) cell lysate <br />Lane 7: Pig brain tissue lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 12 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/10,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 92 kDa <br />Observed band size: 94 kDa

    Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (HA751978) at 1/10,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: MCF7 (Human breast cancer cell) cell lysate
    Lane 3: PANC-1 (Human epidermoid carcinoma pancreatic ductal cell) cell lysate
    Lane 4: MEF (Mouse embryonic fibroblast) cell lysate
    Lane 5: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
    Lane 6: COS-1 (African green monkey kidney fibroblast) cell lysate
    Lane 7: Pig brain tissue lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 12 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA751978, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 92 kDa
    Observed band size: 94 kDa

  • Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (<a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>) at 1/5,000 dilution. <br /> <br />Lane 1: Mouse brain tissue lysate <br />Lane 2: Mouse stomach tissue lysate <br />Lane 3: Rat brain tissue lysate <br />Lane 4: Rat stomach tissue lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 8 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 92 kDa <br />Observed band size: 94 kDa

    Western blot analysis of GRP94 on different lysates with Rabbit anti-GRP94 antibody (HA751978) at 1/5,000 dilution.

    Lane 1: Mouse brain tissue lysate
    Lane 2: Mouse stomach tissue lysate
    Lane 3: Rat brain tissue lysate
    Lane 4: Rat stomach tissue lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 8 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA751978, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 92 kDa
    Observed band size: 94 kDa

  • Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Human <br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br /> <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/500, overnight at 4°C. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature. <br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA751978, 1/500, overnight at 4°C.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Mouse <br />Sample: NIH/3T3 (Mouse fibroblast) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br /> <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/2,500, overnight at 4°C. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature. <br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Mouse
    Sample: NIH/3T3 (Mouse fibroblast)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.

    Primary antibody: HA751978, 1/2,500, overnight at 4°C.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Kidney <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Kidney
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Colon <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Colon
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Kidney <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Kidney
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Kidney <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Kidney
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Mouse <br />Tissue: Colon <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Colon
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Rat <br />Tissue: Colon <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Colon
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA751978, 1/5,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Flow Cytometry (Intra) <br /> <br />Species: Human <br />Sample: HeLa (Human cervix adenocarcinoma epithelial cell) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Tween-20, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br /> <br />Primary antibody: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>(1 μg/mL, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature.

    Application: Flow Cytometry (Intra)

    Species: Human
    Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: HA751978(1 μg/mL, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

  • Immunoprecipitation (IP) <br /> <br />GRP94 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. <br /> <br />Lane 1: HeLa cell lysate (input) <br />Lane 2: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a> IP in HeLa cell lysate <br />Lane 3: Rabbit IgG instead of <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a> in HeLa cell lysate <br /> <br />Exposure time: 7 seconds <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Primary dilution: <a href="/products/HA751978" style="font-weight: bold;text-decoration: underline;">HA751978</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature <br /> <br />Predicted band size: 92 kDa <br />Observed band size: 94 kDa

    Immunoprecipitation (IP)

    GRP94 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA751978 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751978 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA751978 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA751978 in HeLa cell lysate

    Exposure time: 7 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA751978, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 92 kDa
    Observed band size: 94 kDa

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