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Anti-SMARCC1/BAF155 Rabbit Antibody [E10L22]

Catalog No.: F3789

Application: Reactivity: Rat, Human

Experiment Essentials

WB
Recommended SDS-PAGE separating gel concentration: 5%.

Usage Information

Dilution
WB1:1000- 1:10000 IP1:10 - 1:100 IF1:100 FCM1:10 - 1:100

Application
WB, IP, IF, FCM, ChIP

Reactivity
Rat, Human

Source
Rabbit

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
123 kDa 112 kDa, 155 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Rat testis; HeLa; HEK293; Jurkat; K562; 293T; MDA-MB-231
Negative Control

Exprimental 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. 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) and put the sample on ice for 5 min. 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) and put the sample on ice for 5 min. 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: 5%. 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 5% skim milk powder 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.

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.

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) and put the sample on ice for 5 min.

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) and put the sample on ice for 5 min.

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: 5%. 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 5% skim milk powder 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.

IF
Experimental Protocol： Specimen Preparation 1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% Paraformaldehyde diluted in 1X PBS. NOTE: Paraformaldehyde is toxic, use only in a fume hood. 2. Fix cells for 15 min at room temperature. 3. Aspirate fixative, rinse three times in 1X PBS for 5 min each. 4. Proceed with Immunostaining. Immunostaining 1. Add theblocking buffer and incubate for 60 min at RT. 2. Prepare primary antibody diluent in antibody dilution buffer as recommended . 3. Aspirate blocking solution, apply diluted primary antibody. 4. Incubate overnight at 4°C. 5. Rinse three times in 1X PBS for 5 min each. 6. Incubate specimens in fluorochrome-conjugated secondary antibody diluted in antibody dilution buffer for 1–2 hr at room temperature in the dark. 7. Rinse three times in 1X PBS for 5 min each. 8. Mount slides usingmounting medium with DAPI and cover with coverslips. 9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 23°C protected from light.

Experimental Protocol：

Specimen Preparation

1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% Paraformaldehyde diluted in 1X PBS.

NOTE: Paraformaldehyde is toxic, use only in a fume hood.

2. Fix cells for 15 min at room temperature.

3. Aspirate fixative, rinse three times in 1X PBS for 5 min each.

4. Proceed with Immunostaining.

Immunostaining

1. Add theblocking buffer and incubate for 60 min at RT.

2. Prepare primary antibody diluent in antibody dilution buffer as recommended .

3. Aspirate blocking solution, apply diluted primary antibody.

4. Incubate overnight at 4°C.

5. Rinse three times in 1X PBS for 5 min each.

6. Incubate specimens in fluorochrome-conjugated secondary antibody diluted in antibody dilution buffer for 1–2 hr at room temperature in the dark.

7. Rinse three times in 1X PBS for 5 min each.

8. Mount slides usingmounting medium with DAPI and cover with coverslips.

9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 23°C protected from light.

Datasheet & SDS

Biological Description

Specificity
SMARCC1/BAF155 (E10L22) Rabbit mAb detects endogenous levels of total SMARCC1/BAF155 protein.

Subcellular Location
Cytoplasm, Nucleus

Uniprot ID
Q92922

Clone
E10L22

Synonym(s)
BAF155, SMARCC1, SWI/SNF complex subunit SMARCC1, BRG1-associated factor 155, SWI/SNF complex 155 kDa subunit, SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily C member 1

Background
The mSWI/SNF, also known as BAF complex, is an ATP-dependent chromatin remodeling system built from 16 protein subunits encoded by 31 different genes. These subunits assemble in various combinations to form distinct complexes with specialized roles in key biological processes. One universally present core component is BAF155 (SMARCC1), which can exist as a homodimer or as a heterodimer with its paralog BAF170 (SMARCC2). In stem cells, BAF155 is the predominant form, while BAF170 becomes more abundant during cellular differentiation. BAF155 is unique in being the only shared subunit of the embryonic stem cell–specific SWI/SNF complex (esBAF) and the non-canonical BAF (ncBAF) complex. This protein has been implicated as a therapeutic target for colorectal and prostate cancers, and mutations in BAF155 are associated with particularly poor cancer outcomes. The SMARCC1 gene, located on chromosome 3, encodes BAF155. As a member of the SWI/SNF chromatin remodeling family, SMARCC1-containing proteins exhibit helicase and ATPase-related functions, enabling them to regulate transcription by restructuring chromatin around target genes. BAF155 contains a leucine zipper motif typical of transcription-associated proteins and interacts with multiple partners including BAZ1B, ING1, SIN3A, SMARCA2, SMARCA4, and SMARCB1. Functionally, SMARCC1 modulates nucleosome architecture by stimulating the ATPase activity of the complex’s catalytic subunit, thereby influencing gene expression. It is a component of both the neural progenitor–specific BAF (npBAF) complex and the neuron-specific BAF (nBAF) complex. During neural development, neurons transition from npBAF to nBAF complexes as they exit the cell cycle and mature. This subunit switch replaces ACTL6A/BAF53A and PHF10/BAF45A in npBAF with ACTL6B/BAF53B and either DPF1/BAF45B or DPF3/BAF45C in nBAF. The npBAF complex supports neural stem cell self-renewal and proliferation, while nBAF, often working in partnership with CREST, regulates the transcription of genes involved in dendritic growth.

Background

The mSWI/SNF, also known as BAF complex, is an ATP-dependent chromatin remodeling system built from 16 protein subunits encoded by 31 different genes. These subunits assemble in various combinations to form distinct complexes with specialized roles in key biological processes. One universally present core component is BAF155 (SMARCC1), which can exist as a homodimer or as a heterodimer with its paralog BAF170 (SMARCC2). In stem cells, BAF155 is the predominant form, while BAF170 becomes more abundant during cellular differentiation. BAF155 is unique in being the only shared subunit of the embryonic stem cell–specific SWI/SNF complex (esBAF) and the non-canonical BAF (ncBAF) complex. This protein has been implicated as a therapeutic target for colorectal and prostate cancers, and mutations in BAF155 are associated with particularly poor cancer outcomes. The SMARCC1 gene, located on chromosome 3, encodes BAF155. As a member of the SWI/SNF chromatin remodeling family, SMARCC1-containing proteins exhibit helicase and ATPase-related functions, enabling them to regulate transcription by restructuring chromatin around target genes. BAF155 contains a leucine zipper motif typical of transcription-associated proteins and interacts with multiple partners including BAZ1B, ING1, SIN3A, SMARCA2, SMARCA4, and SMARCB1. Functionally, SMARCC1 modulates nucleosome architecture by stimulating the ATPase activity of the complex’s catalytic subunit, thereby influencing gene expression. It is a component of both the neural progenitor–specific BAF (npBAF) complex and the neuron-specific BAF (nBAF) complex. During neural development, neurons transition from npBAF to nBAF complexes as they exit the cell cycle and mature. This subunit switch replaces ACTL6A/BAF53A and PHF10/BAF45A in npBAF with ACTL6B/BAF53B and either DPF1/BAF45B or DPF3/BAF45C in nBAF. The npBAF complex supports neural stem cell self-renewal and proliferation, while nBAF, often working in partnership with CREST, regulates the transcription of genes involved in dendritic growth.

References
https://pubmed.ncbi.nlm.nih.gov/33953332/ https://pubmed.ncbi.nlm.nih.gov/19785031/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

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

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

* Indicates a Required Field

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%