BMP7 Antibody [J15D7] | Primary Antibodies

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:1000 - 1:10000 IP1:150

Application
WB, IP

Reactivity
Mouse, Rat, Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
49 kDa 49 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Fetal kidney; Human prostate; Rat kidney; Mouse kidney; MCF7 cells
Negative Control

Experimental Methods

WB
Experimental Protocol: Sample preparation 1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes. 2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes. 4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant; 5. Remove a small volume of lysate to determine the protein concentration; 6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice. Electrophoretic separation 1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations 2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.) Transfer membrane 1. Take out the converter, soak the clip and consumables in the pre-cooled converter; 2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer; 3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip"; 4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications Recommended conditions for wet transfer: 200 mA, 120 min. ( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.) Block 1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes; 2. Incubate the film in the blocking solution for 1 hour at room temperature; 3. Wash the film with TBST for 3 times, 5 minutes each time. Antibody incubation 1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:1000), gently shake and incubate with the film at 4°C overnight; 2. Wash the film with TBST 3 times, 5 minutes each time; 3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour; 4. After incubation, wash the film with TBST 3 times for 5 minutes each time. Antibody staining 1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly; 2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

WB

Experimental Protocol:

Sample preparation

1. Tissue: Lyse the tissue sample by adding an appropriate volume of ice-cold RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail)，and homogenize the tissue at a low temperature or lyse it by sonication on ice, then incubate on ice for 30 minutes.

2. Adherent cell: Aspirate the culture medium and wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

3. Suspension cell: Transfer the culture medium to a pre-cooled centrifuge tube. Centrifuge and aspirate the supernatant. Wash the cells with ice-cold PBS twice. Lyse the cells by adding an appropriate volume of RIPA/NP-40 Lysis Buffer (containing Protease Inhibitor Cocktail) , sonicate to lyse the cells, and incubate on ice for 30 minutes.

4. Place the lysate into a pre-cooled microcentrifuge tube. Centrifuge at 4°C for 15 min. Collect the supernatant;

5. Remove a small volume of lysate to determine the protein concentration;

6. Combine the lysate with protein loading buffer. Boil 20 µL sample under 95-100°C for 5 min. Centrifuge for 5 min after cool down on ice.

Electrophoretic separation

1. According to the concentration of extracted protein, load appropriate amount of protein sample and marker onto SDS-PAGE gels for electrophoresis. Recommended separating gel (lower gel) concentration: 10%. Reference Table for Selecting SDS-PAGE Separation Gel Concentrations

2. Power up 80V for 30 minutes. Then the power supply is adjusted (110 V~150 V), the Marker is observed, and the electrophoresis can be stopped when the indicator band of the predyed protein Marker where the protein is located is properly separated. (Note that the current should not be too large when electrophoresis, too large current (more than 150 mA) will cause the temperature to rise, affecting the result of running glue. If high currents cannot be avoided, an ice bath can be used to cool the bath.)

Transfer membrane

1. Take out the converter, soak the clip and consumables in the pre-cooled converter;

2. Activate PVDF membrane with methanol for 1 min and rinse with transfer buffer;

3. Install it in the order of "black edge of clip - sponge - filter paper - filter paper - glue -PVDF membrane - filter paper - filter paper - sponge - white edge of clip";

4. The protein was electrotransferred to PVDF membrane. ( 0.45 µm PVDF membrane is recommended ) Reference Table for Selecting PVDF Membrane Pore Size Specifications

Recommended conditions for wet transfer: 200 mA, 120 min.

( Note that the transfer conditions can be adjusted according to the protein size. For high-molecular-weight proteins, a higher current and longer transfer time are recommended. However, ensure that the transfer tank remains at a low temperature to prevent gel melting.)

Block

1. After electrotransfer, wash the film with TBST at room temperature for 5 minutes;

2. Incubate the film in the blocking solution for 1 hour at room temperature;

3. Wash the film with TBST for 3 times, 5 minutes each time.

Antibody incubation

1. Use primary antibody dilution buffer to prepare the primary antibody working liquid (recommended dilution ratio for primary antibody 1:1000), gently shake and incubate with the film at 4°C overnight;

2. Wash the film with TBST 3 times, 5 minutes each time;

3. Add the secondary antibody to the blocking solution and incubate with the film gently at room temperature for 1 hour;

4. After incubation, wash the film with TBST 3 times for 5 minutes each time.

Antibody staining

1. Add the prepared ECL luminescent substrate (or select other color developing substrate according to the second antibody) and mix evenly;

2. Incubate with the film for 1 minute, remove excess substrate (keep the film moist), wrap with plastic film, and expose in the imaging system.

Datasheet & SDS

Biological Description

Specificity
BMP7 Antibody (Rabbit mAb) [J15D7] detects endogenous levels of total BMP7 protein.

Subcellular Location
Secreted

Uniprot ID
P18075

Clone
J15D7

Synonym(s)
OP1 \| BMP7 \| Bone morphogenetic protein 7 \| BMP-7 \| Osteogenic protein 1 \| OP-1

Background
BMP7, also known as osteogenic protein-1, belongs to the TGF-β superfamily of secreted growth factors that orchestrate tissue morphogenesis, homeostasis, and repair across skeletal, renal, and neural systems. Synthesized as inactive pro-BMP7, it undergoes furin-mediated cleavage to yield the mature disulfide-linked homodimer with a characteristic TGF-β fold featuring wrist and knuckle epitopes for receptor engagement. BMP7 binds preformed heterotetrameric complexes of type I (ALK2/3/6) and type II (BMPRII/ActRIIA/B) serine/threonine kinase receptors; type II phosphorylates type I GS domain, activating its kinase to propagate canonical Smad1/5/8 phosphorylation, complex formation with Smad4, and nuclear translocation to drive target genes like ID1, Runx2, and Msx2 for osteoblast/chondrocyte differentiation. Non-canonical arms engage TAK1-TAB1/2/3 for p38/ERK/JNK MAPKs promoting proliferation/migration, or PI3K-Akt-mTOR inhibiting apoptosis and inflammation. Antagonists like noggin, chordin, gremlin, and follistatin sequester BMP7 extracellularly, while intracellularly Smurf1/2 ubiquitinates receptors/Smads for degradation, and Smad6/7 competitively inhibit R-Smad phosphorylation. Physiologically, BMP7 patterns kidney metanephric mesenchyme via ureteric bud branching and nephron progenitor proliferation, maintains renal epithelial integrity against fibrosis, supports brown adipogenesis, and regulates monocyte polarization toward anti-inflammatory M2 macrophages. TGF-β/Smad3 upregulates Smurf2 to suppress BMP7/Smad1/5/8 while BMP7 reciprocally attenuates TGF-β/Smad3 via miR-29/miR-200 induction countering miR-21/192-driven fibrosis. BMP7 loss exacerbates epithelial-mesenchymal transition, leading to tubulointerstitial scarring; recombinant BMP7 restores Smad balance, reverses EMT, and protects against diabetic nephropathy progression.

Background

BMP7, also known as osteogenic protein-1, belongs to the TGF-β superfamily of secreted growth factors that orchestrate tissue morphogenesis, homeostasis, and repair across skeletal, renal, and neural systems. Synthesized as inactive pro-BMP7, it undergoes furin-mediated cleavage to yield the mature disulfide-linked homodimer with a characteristic TGF-β fold featuring wrist and knuckle epitopes for receptor engagement. BMP7 binds preformed heterotetrameric complexes of type I (ALK2/3/6) and type II (BMPRII/ActRIIA/B) serine/threonine kinase receptors; type II phosphorylates type I GS domain, activating its kinase to propagate canonical Smad1/5/8 phosphorylation, complex formation with Smad4, and nuclear translocation to drive target genes like ID1, Runx2, and Msx2 for osteoblast/chondrocyte differentiation. Non-canonical arms engage TAK1-TAB1/2/3 for p38/ERK/JNK MAPKs promoting proliferation/migration, or PI3K-Akt-mTOR inhibiting apoptosis and inflammation. Antagonists like noggin, chordin, gremlin, and follistatin sequester BMP7 extracellularly, while intracellularly Smurf1/2 ubiquitinates receptors/Smads for degradation, and Smad6/7 competitively inhibit R-Smad phosphorylation. Physiologically, BMP7 patterns kidney metanephric mesenchyme via ureteric bud branching and nephron progenitor proliferation, maintains renal epithelial integrity against fibrosis, supports brown adipogenesis, and regulates monocyte polarization toward anti-inflammatory M2 macrophages. TGF-β/Smad3 upregulates Smurf2 to suppress BMP7/Smad1/5/8 while BMP7 reciprocally attenuates TGF-β/Smad3 via miR-29/miR-200 induction countering miR-21/192-driven fibrosis. BMP7 loss exacerbates epithelial-mesenchymal transition, leading to tubulointerstitial scarring; recombinant BMP7 restores Smad balance, reverses EMT, and protects against diabetic nephropathy progression.

References
https://pubmed.ncbi.nlm.nih.gov/31979268/ https://pubmed.ncbi.nlm.nih.gov/23126427/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%

BMP7 Antibody (Rabbit mAb) [J15D7]

Usage Information

Experimental Methods

Datasheet & SDS

Biological Description

Tech Support